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Freshwater network ecosystems consist of interconnected lotic and lentic environments within the same catchment area. Using Plitvice Lakes as an example, we studied the changes in environmental conditions and microbial communities (bacteria and fungi) that occur with downstream flow. Water samples from tributaries, interlake streams, connections of the cascading lakes, and the Korana River, the main outflow of the system, were characterized using amplicon sequencing of bacterial 16S rRNA and fungal ITS2 genes. Our results show that different environmental conditions and bacterial and fungal communities prevail among the three stream types within the freshwater network ecosystem during multiple sampling seasons. Microbial community differences were also confirmed along the longitudinal gradient between the most distant sampling sites. The higher impact of “mass effect” was evident during spring and winter, while “species sorting” and “environmental selection” was more pronounced during summer. Prokaryotic community assembly was majorly influenced by deterministic processes, while fungal community assembly was highly dominated by stochastic processes, more precisely by the undominated fraction, which is not dominated by any process. Despite the differences between stream types, the microbial community of Plitvice Lakes is shown to be very stable by the core microbiome that makes up the majority of stream communities. Our results suggest microbial community succession along the river-lake continuum of microbial communities in small freshwater network ecosystems with developed tufa barriers.

Purpose In aquatic ecosystems, protists play a crucial role and cover numerous ecological functions. The karstic Krka River (Croatia) is a unique hotspot for high diversity of aquatic organisms, especially protists. The main objective of the present study was to obtain a detailed overview of the protist community structure in the periphyton of the Krka River and to determine the differences in protist diversity along the river. Methods Protist diversity was detected by amplicon sequencing of the hypervariable region V9 of the 18S rRNA gene, using the universal eukaryotic primer pair. Results The three main groups of protists were as follows: Ciliophora, Cercozoa, and Bacillariophyta. In terms of abundance of protist OTUs, the shade plot revealed an evident difference from the upstream to downstream river section, which increased between locations from Krka spring to Skradinski buk. Diversity was explored using measures of alpha and beta diversity. Alpha diversity showed an increasing trend in the downstream direction of the river. The location effect, or clustering/grouping of samples by location, was confirmed by the PERMANOVA permutation test of beta diversity. Conclusion The combination of alpha and beta diversity can help provide deeper insight into the study of diversity patterns, but also point out to decline in species diversity and allow for effective ways to protect aquatic karst habitats in future management.

In this preliminary study, we applied the Multivariate Curve Resolution- Alternating Least Squares (MCR-ALS) method to analyze the excitation-emission matrices of multifloral honey samples, combined with differential scanning calorimetry (DSC) to estimate infection of honey bee colonies with N. ceranae or V. destructor. Fluorescence spectroscopy combined with MCR-ALS was used to determine the ratio of the spectral components originating from proteins (C1) and phenolics (C2), two minor but essential constituents of honey, as a ratiometric indicator of infection level in related hives. The C1/C2 ratio decreased linearly with the increase of infection in both N. ceranae and V. destructor cases, the R2 was 0.941 and 0.912, respectively. Additionally, DSC has shown that the magnitude of changes in sugar environments of the honey samples, reflected in sugar phase transitions, rises with increasing infection level in bee colonies. These results indicate that fluorescence combined with MCR-ALS could be used for rapid, non-destructive and cheap screening of honey to estimate the level of infection of honey bee colonies.

Leaf shape in European beech provenances varies geographically, with narrower and longer lamina observed in southern provenances, indicating a direct selection favoring leaf shape that likely safeguards trees performances under less favorable growing conditions. Spatial and environmental patterns of European beech (Fagus sylvatica L.) leaf size and shape variations were studied using landmark-based geometric morphometrics. The study involved eight provenances originating from three biogeographic regions (i.e., Alpine, Continental, and Pannonian), spanning across six European countries (Bosnia and Herzegovina, Croatia, Germany, Hungary, Romania and Serbia). All specimens were cultivated in a common garden experiment. The symmetric component of leaf shape variation was analyzed by Procrustes ANOVA and multi‐variate analyses [principal component analysis and canonical variate analysis (CVA)], whereas MANOVA was used to examine asymmetry. Partial least square (PLS) analysis was used to assess the covariation between leaf shape and size, and geographical position and environmental variables at the sites of provenance origin, respectively. A highly observed phenotypic variation for the shape and size of leaf both within and among provenances, indicates a strong local adaptation of provenances within the species natural range. CVA revealed the existence of two clusters of provenances based on the leaf shape, i.e., the first group included provenances originating from Balkan Peninsula and Central Europe, while the second group consisted of two Romanian provenances and the northernmost provenance from Germany. Likewise, PLS evidenced that leaf shape was spatially structured along latitudinal (− 0.64) and longitudinal (0.60) gradients, with southern provenances having longer and narrower (ovate) lamina compared to the northern ones. In contrast, no correlation was found between centroid size and spatial and environmental variables. Results suggest the presence of direct selection favoring a leaf shape that likely safeguards trees from heat and water loses under less favorable growing conditions.

Background Monolithic chromatography using convective interaction media (CIM) disks or columns can be used in the separation step of speciation analysis. When different monolithic disks are placed in one housing, forming conjoint liquid chromatography (CLC) monolithic column, two-dimensional separation is achieved in a single chromatographic run. Methods Here, we assembled low-pressure (maximum 50 bar) CLC monolithic column, which consists of two 0.34 mL shallow CIM monolithic disks and high-pressure CLC column (maximum 150 bar) from 0.1 mL analytical high performance short bed CIMac monolithic disks. Both the CLC columns constructed from affinity Protein G and weak anion exchange diethylamine (DEAE) disks, were applied for the speciation of cisplatin, oxaliplatin and carboplatin in spiked standard serum proteins, spiked human serum and serum of cancer patients. The analytical performances of the CLC columns used were evaluated by comparing their robustness, selectivity, repeatability and reproducibility. The separated serum proteins were detected on-line by ultraviolet (UV) and eluted Pt species by inductively coupled plasma mass spectrometry (ICP-MS). For accurate quantification of the separated Pt species (unbound Pt-based chemotherapeutic from species associated to transferrin (Tf), human serum albumin (HSA) and Immunoglobulin G (IgG)), post column isotope dilution (ID)-ICP-MS was used. Results The data from analyses showed that both tested CLC monolithic columns gave statistically comparable results, with the low-pressure CLC column exhibiting better resolving power and robustness. It also enables more effective cleaning of monolithic disks and to analyse larger series of serum samples than the high-pressure CLC column. Analyses of serum samples of cancer patients treated with cisplatin or carboplatin showed that Pt-chemotherapeutics were bound preferentially to HSA (around 80%). The portion of unbound Pt in general did not exceed 2%, up to 5% of Pt was associated with Tf and approximately 20% with IgG. Column recoveries, calculated as a ratio between the sum of concentrations of Pt species eluted and concentration of total Pt in serum samples, were close to 100%. Conclusions Low-pressure CLC column exhibited greater potential than high-pressure CLC column, and can be thus recommended for its intended use in speciation analysis of metal-based biomolecules.

The rodent associations from the late Oligocene deposits of Ugljevik and Paragovo in the Pannonian basin are described. Pannoniamys paragovensis, a new genus and species of Spalacidae, is defined. The published data from a third late Oligocene locality in the area (Banovići) is included in the discussion of the changes in community structure through the time-interval studied. Although the composition of the rodent faunas studied differs, the dominating cricetid genus in all three associations is Deperetomys, a genus that appears in western Europe during the late middle Miocene. While the late Oligocene rodent record of central and southwestern Europe is characterised by endemic development, the regime in the Balkans seems to have been one of multiple immigrations and replacements. The late Oligocene age of the rodent fauna from Ugljevik estimated at about 26 Ma, provides a minimum age for the start of rifting in the Pannonian basin, some 6 Ma earlier than previously documented.

Nutrients play an essential role in many metabolic processes whose deficiency or excess can be harmful to the plant itself and through the food chain to both animals and humans. Medicinal plants used in the food and pharmaceutical industries can be contaminated with increased concentrations of heavy metals. The plant species Seseli rigidum and Seseli pallasii from the Balkan Peninsula are used in traditional medicine and spices in the diet, so it was necessary to determine the mineral composition to ensure their safe application. In this work, the mineral composition was determined in medicinal species of the genus Seseli using inductively coupled plasma with optical emission spectrometry (ICP-OES). Two multivariate statistic methods –principal component analysis (PCA) and hierarchical cluster analysis (HCA) were applied to distinguish samples regarding their mineral composition. The mineral composition of both studied species is following the literature data. The results obtained using multivariate statistics methods agree and distinguish certain parts of the tested plants based on the highest content of micro, macro, or trace elements.

Heat stress is one of the most important environmental factors that influences wheat growth and development, leading to significant losses in grain yield and has become a significant detrimental factor for worldwide wheat production. In recent years, several studies suggested that eukaryotic elongation factor 1A (eEF1A), may contribute to heat tolerance in plants, therefore the aim of this study was: to investigate the accumulation of eEF1A in wheat under conditions of moderate and high air temperatures; to determine the amount of photosynthetic pigments and to determine the yield traits; and to examine whether there is a correlation between eEF1A accumulation, photosynthetic pigments, and yield in different wheat varieties. The results showed that heat stress induced accumulation of eEF1A significantly different among wheat varieties and showed that varieties with a higher accumulation of eEF1A under heat stress are characterized by a smaller decrease in the photosynthetic pigments. A correlation between higher accumulation of eEF1A under heat stress and yield traits was found. Analyzed parameters from two growing seasons, indicated that the higher accumulation of eEF1A and a smaller decrease in photosynthetic pigments distinguishes the varieties more resistant to heat stress. The analysis of the molecular mechanisms by immunoblot, under conditions of high and moderate air temperatures in two growing seasons, aims to develop agricultural strategy and develop wheat varieties tolerant to heat stress.

We present a checklist of 86 valid freshwater mollusc species of Serbia (65 gastropods and 21 bivalves) belonging to 19 families (15 gastropods and four bivalves). The list is based on our latest research and includes data from published sources and personal communications. The most diverse family among gastropods is Planorbidae (16 species), while Sphaeriidae are most diverse within the bivalves (10 species). Ten species are local endemics (all of them gastropods), while eight are introduced species (three gastropods and five bivalves). The Danube River and its smaller tributaries harbour the most diverse gastropod community, with a total of 61 species (41 gastropods and 20 bivalves). The snail Theodoxus transversalis (Pfeiffer, 1828) and the bivalve Unio crassus Philipson, 1788 are listed as EN (endangered) on the global (IUCN) level, while the snail Plagigeyeria gladilini Kuščer, 1937 and the bivalves Pseudanodonta complanata (Rossmässler, 1835) and Sphaerium rivicola (Lamarck, 1818) are considered as VU (vulnerable) species. Of those IUCN taxa, only U. crassus is listed as protected on the national level (along with 12 mostly local endemic gastropod species). In comparison with the previous checklist of gastropods by Karaman & Karaman (2007), a total of seven species are added here: Belgrandiella serbica Glöer 2008, Bythinella istoka Glöer & Pešić 2014, B. nonveilleri Glöer 2008, B. pesterica Glöer 2008, Acroloxus lacustris (Linnaeus, 1758), Melanoides tuberculata (O.F. Müller, 1774) and Armiger crista (Linnaeus, 1758). In view of global and regional trends in taxonomy, ecology and invasive biology, an even richer diversity of the Serbian freshwater malocofauna can be expected.

Hereditary spastic paraplegia (HSP) is among the most genetically diverse of all monogenic diseases. The aim was to analyze the genetic causes of HSP among adult Serbian patients. The study comprised 74 patients from 65 families clinically diagnosed with HSP during a nine-year prospective period. A panel of thirteen genes was analyzed: L1CAM (SPG1), PLP1 (SPG2), ATL1 (SPG3A), SPAST (SPG4), CYP7B1 (SPG5A), SPG7 (SPG7), KIF5A (SPG10), SPG11 (SPG11), ZYFVE26 (SPG15), REEP1 (SPG31), ATP13A2 (SPG78), DYNC1H1, and BICD2 using a next generation sequencing-based technique. A copy number variation (CNV) test for SPAST, SPG7, and SPG11 was also performed. Twenty-three patients from 19 families (29.2%) had conclusive genetic findings, including 75.0% of families with autosomal dominant and 25.0% with autosomal recessive inheritance, and 15.7% of sporadic cases. Twelve families had mutations in the SPAST gene, usually with a pure HSP phenotype. Three sporadic patients had conclusive findings in the SPG11 gene. Two unrelated patients carried a homozygous pathogenic mutation c.233T>A (p.L78*) in SPG7 that is a founder Roma mutation. One patient had a heterozygous de novo variant in the KIF5A gene, and one had a compound heterozygous mutation in the ZYFVE26 gene. The combined genetic yield of our gene panel and CNV analysis for HSP was around 30%. Our findings broaden the knowledge on the genetic epidemiology of HSP, with implications for molecular diagnostics in this region.

The content of daidzein, genistein, formononetin, and biochanin A isoflavones was studied in natural populations of red and Hungarian clover, to estimate their impact on fodder quality and to determine directions in possible breeding programs. The study included 6 red clover (Trifolium pratense) and 6 Hungarian clover (Trifolium pannonicum) populations, collected in the central Balkans. The differences between the species and among the populations were analysed. The average content of total isoflavones was 1.393 mg g–1 and 0.487 mg g–1 of air dry matter in Hungarian clover, respectively. While the most prevailed isoflavone in red clover was biochanin A (46%), the Hungarian clover populations were rich in genistein (43%). The red clover leaves accumulated the highest content of isoflavones. The Hungarian clover flowers and leaves had an equal amount of isoflavones. The obtained values of the total isoflavones could not affect the overall nutrient quality and therefore, researched natural populations of two clover species could be considered for further breeding programs.

Within the nursery sector, a global interest in functional fruits imposes a permanent need to improve the tissue culture technology. A modern bioreactor immersion system is currently a valid solution for the micropropagation of many cultures. At the Faculty of Agrobiotechnical Sciences Osijek (FAZOS, Croatia), a research was conducted with the aim of improving the raspberry (Rubus idaeus L., cv. HimboTop®) micropropagation protocol. The treatments implied the modifications of the MS and DKW nutrient medium (DKW – standard; DKWm – substitution of FeEDTA with the FeEDDHA; MSm – macroelement modification, 3 x CaCl2, MgSO4, and KH2PO4 + FeEDDHA), differently depending on the aggregate condition (semi‐solid and liquid SETISTM). Subsequent to 15 (SETISTM) and 30 cultivation days (semi-solid medium), respectively, a morphological parameter analysis and a multiplication success evaluation were commenced. In all treatments (DKW, DKWm, and MSm), the observed parameters (shoot height, number of leaves, leaf width and length) were significantly better in an interaction with the liquid medium. Comparing the models, a liquid medium (SETISTM) resulted in a significantly higher explant multiplication (4.02) (a semi‐solid medium - 2.74). The best reproduction quality, implying a better plant vigor with a reduced vitrification, was obtained on the MSm medium in the SETISTM. A concentration optimization of individual mesocomponents (i.e., of the macroelements) and their chemical forms (FeEDDHA) may contribute to their improved adoption and consequently to the stress suppression due to explant transplantation. Selecting the modern technical solutions (TIS bioreactors - SETISTM) and continuously improving the protocols, we may obtain the realistically sustainable models, suitable for a massive clonal reproduction.

Blackleg outbreaks were noticed on three fields (about 100 ha total) in 2 consecutive years (2018, 2019) in one of the main potato growing areas in Serbia (Bačka region, Vojvodina). The percentage of infected plants reached 40 to 70%, with 10.5 to 44.7% yield reductions. From the three fields, out of 90 samples Pectobacterium carotovorum subsp. brasiliensis was most frequently identified and diagnosed as causal agent of potato blackleg in Serbia for the first time (29 isolates). Dickeya dianthicola was a less frequently causative bacterium, which was also noticed for the first time (nine isolates). A total of 38 isolates were characterized based on their phenotypic and genetic features, including a pathogenicity test on potato. The repetitive element PCR (rep-PCR) using BOX, REP, and ERIC primer pairs differentiated five genetic profiles among 38 tested isolates. Multilocus sequence analysis (MLSA) of four housekeeping genes, acnA, gapA, icdA, and mdh, revealed the presence of three so far unknown P. c. subsp. brasiliensis multilocus genotypes and confirmed clustering into two main genetic clades as determined in other studies. MLSA also revealed the presence of a new genotype of D. dianthicola in Serbia.

This paper provides essential information on medicinal plant uses in the veterinary ethnopharmacology of Pirot County in Eastern Serbia, known as a relatively isolated, multiethnic, and a traditional agricultural area. The aim of the study was to collect, analyze, and evaluate the medicinal plants' ethnoveterinary knowledge in a Pirot County (2761 km2, total 92,479 inhabitants, of which 34,672 in the rural area). The local inhabitants of the rural area were interviewed by a semi-structured questionnaire to determine how many of them are familiar with the plants' application in veterinary ethnopharmacology. Interviews were conducted in four municipalities of the Pirot County, where the samples of the local rural population included 631 respondents (mean age 50; 45.59% women, 53.41% men) at 144 sites, of which 148 respondents from 92 villages reported the use of plants in veterinary medicine. The data are presented in a tabular manner and include scientific and local names, the part used, pharmaceutical form, therapeutic indication, and mode of administration, followed with several reports. Furthermore, most frequently reported taxa are systemized for each therapeutic group municipality-wise with calculated informant consensus factor. A comparison with previously published data collected from surrounding territories on the Balkan Peninsula was performed by considering the Jaccard index. The survey revealed 192 plants used by the local population for various health indications, of which 46 species were reported for use in veterinary medicine. Plants reported for the treatment of health issues in livestock (206 reports) were distributed in 30 families out of which the main ones being Asteraceae (8 species), Lamiaceae (4 species), Rosaceae (4 species) and the most abundant were Asteraceae (43 reports), Hypericaceae (28 reports), Polygonaceae (26 reports) and Ranunculaceae (23 reports). The most frequently used plants in veterinary ethnopharmacology were Cichorium intybus, Hypericum perforatum, and Rumex patientia. Aerial plant parts were exploited more frequently than underground parts. The mode of administration was primarily oral in the water extract form. The primary reported therapeutic groups were diarrhea (98 reports), wounds (27 reports), dermatological diseases (scabies and pediculosis) (14 reports), and elevated temperature (10 reports). The use of 22 plant species was not reported in other ethnoveterinary surveys in the Balkan Peninsula. Knowledge of plants for veterinary use is still constrained among the population of the rural parts of Pirot County. The study identified plant species used by the local population to deal with health issues in domestic animals. The presented findings of this study can be a good starting point for new phytopharmacological investigations in the veterinary domain.

Cross-talk between phytohormones and sugars is intensely involved in plant metabolism, growth and regeneration. We documented alterations in cytokinin (CK) homeostasis in four developmental stages during de novo shoot organogenesis (DNSO) of kohlrabi (Brassica oleracea var. gongylodes cv. Vienna Purple) seedlings induced by exogenous CKs, trans-zeatin (transZ) and thidiazuron (TDZ), added together with elevated sucrose concentration (6% and 9%). Significant impact of CK and sucrose treatment and their interaction was recorded in all investigated stages, including plantlet development before calli formation (T1 and T2), calli formation (T3) and shoot regeneration (T4). Results showed remarkable increase in total CK levels for transZ treatment, particularly with 9% sucrose. This trend was observed for all physiological and structural groups of CKs. Application of TDZ contributed to little or no increase in CK levels regardless of sucrose concentration. Analysis of expression profiles of organogenesis-related genes involved in auxin transport, CK response, shoot apical meristem formation and cell division revealed that higher sugar concentration significantly downregulated the analysed genes, particularly in T3. This continued on TDZ, but transZ induced an opposite effect with 9% sucrose in T4, increasing gene activity. Our results demonstrated that phytohormone metabolism might be triggered by sucrose signalling in kohlrabi DNSO.

The aim of the present study was to determine and elaborate on all changes in old-aged (OA) versus young-aged (YA) rat thyroids by using stereological, ultrastructural, hormonal, and gene expression analyses. We used 4- and 24-month-old male Wistar rats in our evaluation, presenting all changes in comparison with YA rats. Results showed that the thyroid parenchyma was characterized by higher absolute volumes of the gland, colloid, epithelium, and interstitium by 135, 135, 140, and 142% (p < 0.05) respectively, while the relative volumes of colloid and glands were unchanged. Ultrastructural analysis revealed less active glands, with smaller amounts of lysosomes, thyroglobulin (Tg) granules, and microvilli in the luminal colloid. Optical density values for thyroid peroxidase (TPO), Tg, and vascular-endothelial growth factor immunostaining remained unchanged; however, TPO and Tg exhibited visually stronger expression in small active follicles. Thyroxine (T4)-Tg, the relative intensity of fluorescence (RIF), serum T4, and the sodium-iodide symporter immunohistochemical and gene expressions decreased by 20, 40, 29, and 31% (p < 0.05), respectively, in OA thyroids. Pituitary thyroid-stimulating hormone (TSH) RIF increased by 44% (p < 0.05), but the TSH serum concentration remained unchanged. In conclusion, the obtained results indicate depression of the thyroid gland synthetic and secretory capacity with advanced age.

Hazelnut (Corylus avellana L.) is an economically very important fruit species cultivated in many countries. The world's largest producer and exporter of hazelnuts is Turkey. In Serbia, compared to other types of fruit, hazelnuts are grown on significantly smaller areas. In the last decade, hazelnut has become more interesting for the producers and there is a growing trend of increase of areas under this crop. Hazelnut is the host of numerous pathogens and pests that can cause significant economic damage in orchards. To date, there has been no comprehensive research on the causes of hazelnut diseases in Serbia, and the goal of our work was to determine the causes of some fungal diseases. In the period from 2017 to 2020, samples of hazelnut leaves and branches with symptoms of necrotic spotting were collected from a large number of plantations in Serbia. A large number of isolates were isolated from the collected samples for further determination. Based on the morphological and pathogenic properties of the isolates, the presence of phytopathogenic fungi from the genus Pestalotiopsis was determined.

Bacterial canker and leaf spot caused by plant pathogenic bacterium Pseudomonas is among the most destructive cherry diseases worldwide. Nowadays in Serbia, sweet cherry production significantly increased and the new plantations, mainly grown from imported planting material are being raised every year. During spring, 2018 and 2019, occurrence of bacterial canker and leaf spot symptoms was observed on a newly planted sweet cherry plantations in two localities, Zitoradja (Southern region) and Karavukovo (Northern region-Vojvodina). Typical P. syringae colonies were isolated on Nutrient Sucrose Agar supplemented with 5% sucrose (NSA). A total of fifteen isolates were selected and identified. Results of the LOPAT test (+---+) determined them to belong to fluorescent Pseudomonas Group Ia, while results of G+A+T-Ta- tests indicate presence of Pseudomonas syringae pv. syringae. Pathogenicity was confirmed on immature sweet and sour cherry fruitlets by forming of black, sunken lesions for all tested isolates. Genes syrB and syrD were successfully detected in all tested isolates. DNA sequencing using gapA, gltA, gyrB and rpoD housekeeping genes determined tested isolates to belong to P. s. pv. syringae using the National Center for Biotechnology Information (NCBI) nucleotide BLAST. The Serbian isolates shared 99.47% to 100% (Zitoradja) and 99.38% to 100% (Karavukovo) identity with bacterium P. s. pv. syringae. Phylogenetic analysis grouped isolates from Zitoradja in one tree cluster, separate from the Karavukovo isolates, indicating presence of two genetically diverse groups of causal pathogen P. s. pv. syringae, obtained from two geographically distinct localities in Serbia. Phylogeographic analysis grouped isolates from Zitoradja in multilocus haplotype coded as REz and isolates originated from Karavukovo in multilocus haplotype coded as REk. Considering that during last few years P. syringae continuously occurs mainly in young sweet cherry plantations, where imported material is used for raising, health status check is recommended to be included as obligatory measure when nursery material is used from import.

Rosa canina L. (dog rose) is an important ornamental, edible and medicinal plant. It has been used as a rootstock for ornamental roses, grown in plantations for fruit harvesting and it is suitable for revegetation of abandoned mine lands. The propagation of native genotypes that are well adapted to local conditions can provide planting material for both revegetation and plantation purpose. Micropropagation is the most suitable method for a rapid vegatative propagation of selected wild genotypes, but an increased presence of pathogens as well as higher contamination rate during culture establishment were expected. An occurrence of a specific Fe-chlorosis during in vitro propagation of roses is also possible. Therefore, the optimal period and disinfection protocol for establishing sterile in vitro culture of selected genotypes of dog rose was investigated, as well as an effect of increasing the FeEDTA concentration in the MS medium during multiplication phase. The obtained results showed that the optimal time for taking initial explants corresponds to optimal time for taking green cuttings in traditional vegetative propagation by softwood cuttings, and the best results were achieved using shoots collected in the first week of May, when the flowers were open. The iron chelate concentration in the medium affected the mean number of shoots, and doubling of its concentration resulted in a considerably higher number of shoots per explant.

The aim of this study was to determine the influence of the origin and size of planting material on the morphological characteristics of seed tuber's three potato cultivars. The objects of research were three varieties of potatoes: Aladdin, Newton and Desiree. The analysis of the morphological characteristics tuber showed highly significant (r

This study aimed to investigate the climatological impacts of air temperature, sunshine duration, and rain precipitation, in a moderate climate, on the reproductive performance of Romanov sheep in the continental part of Croatia, during two consecutive years (2019-2020). Our study included 549 matings with 520 successful conceptions at 5 Romanov sheep farms, and the fertility rate was 94.72%. The percentage of lambs delivered in winter was 53.27% (n = 277), in spring 20.19% (n = 105), in summer 14.23% (n = 74) and 12.31% in autumn (n = 64). According to percentile ranks and classification ratings, thermal conditions have been described as very warm (during 2019-2020), while precipitation conditions, expressed in percentiles, were wet (during 2019) but dry (during 2020) for this region. The peak of sexual activity was from mid-August until October in 2019 and from the end of August to November in 2020 for Romanov sheep in northwestern (NW) Croatia. A Kruskal-Wallis test showed a statistically significant difference in the number of pregnant ewes during the different mating seasons in the year (p < 0.001). Therefore, we can assume that air temperature, precipitation volume, and sunshine duration during the different seasons could impact the reproductive performance of Romanov sheep in a moderate climate.

The contribution of domestic cattle in human societies is enormous, making cattle, along with other essential benefits, the economically most important domestic animal in the world today. To expand existing knowledge on cattle domestication and mitogenome diversity, we performed a comprehensive complete mitogenome analysis of the species (802 sequences, 114 breeds). A large sample was collected in Southeast Europe, an important agricultural gateway to Europe during Neolithization and a region rich in cattle biodiversity. We found 1725 polymorphic sites (810 singletons, 853 parsimony-informative sites and 57 indels), 701 unique haplotypes, a haplotype diversity of 0.9995 and a nucleotide diversity of 0.0015. In addition to the dominant T-3 and several rare haplogroups (Q, T-5, T-4, T-2 and T-1), we have identified maternal line in Austrian Murbodner cattle that possess surviving aurochs' mitochondria haplotype P-1 that diverged prior to the Neolithization process. This is convincing evidence for rare female-mediated adaptive introgression of wild aurochs into domesticated cattle in Europe. We revalidated the existing haplogroup classification and provided Bayesian phylogenetic inference with a more precise estimated divergence time than previously available. Occasionally, classification based on partial mitogenomes was not reliable; for example, some individuals with haplogroups P and T-5 were not recognized based on D-loop information. Bayesian skyline plot estimates (median) show that the earliest population growth began before domestication in cattle with haplogroup T-2, followed by Q (-10.0-9.5 kyBP), whereas cattle with T-3 (similar to 7.5 kyBP) and T-1 (similar to 3.0-2.5 kyBP) expanded later. Overall, our results support the existence of interactions between aurochs and cattle during domestication and dispersal of cattle in the past, contribute to the conservation of maternal cattle diversity and enable functional analyses of the surviving aurochs P-1 mitogenome.

Ferulic acid (FA) is used in skin formulations for protection against the damaging actions of the reactive oxygen species (ROS) produced by UVA radiation. Possible underlying protective mechanisms are not fully elucidated. By considering the kinetics of proton-coupled electron transfer (PCET) and radical-radical coupling (RRC) mechanisms, it appears that direct scavenging could be operative, providing that a high local concentration of FA is present at the place of •OH generation. The resulting FA phenoxyl radical, after the scavenging of a second •OH and keto-enol tautomerization of the intermediate, produces 5-hydroxyferulic acid (5OHFA). Inhibition of the lipoxygenase (LOX) enzyme, one of the enzymes that catalyse free radical production, by FA and 5OHFA were analysed. Results of molecular docking calculations indicate favourable binding interactions of FA and 5OHFA with the LOX active site. The exergonicity of chelation reactions of the catalytic Fe2+ ion with FA and 5OHFA indicate the potency of these chelators to prevent the formation of •OH radicals via Fenton-like reactions. The inhibition of the prooxidant LOX enzyme could be more relevant mechanism of skin protection against UVA induced oxidative stress than iron chelation and assumed direct scavenging of ROS.

The newly synthesized coumarin derivative with dopamine, 3-(1-((3,4-dihydroxyphenethyl)amino)ethylidene)-chroman-2,4-dione, was completely structurally characterized by X-ray crystallography. It was shown that several types of hydrogen bonds are present, which additionally stabilize the structure. The compound was tested in vitro against different cell lines, healthy human keratinocyte HaCaT, cervical squamous cell carcinoma SiHa, breast carcinoma MCF7, and hepatocellular carcinoma HepG2. Compared to control, the new derivate showed a stronger effect on both healthy and carcinoma cell lines, with the most prominent effect on the breast carcinoma MCF7 cell line. The molecular docking study, obtained for ten different conformations of the new compound, showed its inhibitory nature against CDKS protein. Lower inhibition constant, relative to one of 4-OH-coumarine, proved stronger and more numerous interactions with CDKS protein. These interactions were carefully examined for both parent molecule and derivative and explained from a structural point of view.

The regeneration of pedunculate oak by shelterwood cutting causes profound ecological changes in the regenerated stands. In this paper, we studied how the pedunculate oak leaf miner community changes after shelterwood cutting with a short regeneration period and does the stand age influence the trend of seasonal variation in their diversity. Leaf miners were chosen because they are a very diverse group of insects closely tied to their host plant. For this reason, their community reacts to all changes to which their host plant is subjected. Three regenerated (5, 10, and 15-year-old) and one control (110-year-old forest) stand were selected in the studied area. A total of 28 species of leaf miners were recorded in them. Many rare species disappeared after shelterwood cutting, while frequent and abundant species experienced a decline in numbers. That led to a decrease in diversity, which was the lowest in the 5-year-old stand. The diversity of leaf miners began to increase in the older regenerated stands. The highest diversity was observed in the control stand. A period of 5 years is needed for the recolonization of approx. 40% of the leaf miner fauna, while 10 years is the recolonization period for approx. 60% and 15 years for approx. 85% of the above fauna. The trend of seasonal variation of the diversity of the pedunculate oak leaf miner community varied in stands of different ages. The diversity of oak leaf miners was the lowest in spring, whereas it increased until the middle of summer and then decreased in all the studied stands.

With the aim of determining the effect of response to heat stress, parity, breed and breeding region on somatic cell count (SCC) in dairy cattle, the individual test-day records of dairy cows collected from 2005 to 2012 in regular milk recording (AT4 / BT4 method) in Croatia were used. The proportional differences in log somatic cell count, and logSCC (in %) in subsequent milk recordings (1st, and 2nd) were determined in cows with a significant decrease in daily milk yield at a set temperature-humidity index, THI threshold (68, 72, and 76). The results indicate high variability in somatic cell count due to the animal’s response to heat stress (heat stressed at THI in 68, 72, and 76), parity (1, 2, and 3+), breed (Holstein, and Simmental) and breeding region (Central, and Eastern Croatia). Also, it could be pointed out that Holsteins, especially cows in higher parities (2, 3+) reared in Eastern Croatia were shown to be more sensitive to environmental stress, even in the period after heat stress (1st, and 2nd subsequent milk recording). Since SCC is not only an indicator for mastitis, but also reflects a cow’s immune response to general stress situations, the high variability determined may be explained by the individual adaptation capabilities of these two breeds to breeding conditions in the analysed regions.

The aim of the study was to investigate genotoxic and cytotoxic activities of Artemisia vulgaris L. and Artemisia alba Turra methanol extracts concerning their chemical composition, separately and in co-treatment with a known mutagen (mitomycin C, MMC). For these evaluations, the cytokinesis-block micronucleus (CBMN) assay as the method for measuring MN frequency in human peripheral blood lymphocytes (PBLs) and MTT assay as the proliferation test in SW-480 human colon cancer cells and human periodontal ligament stem cells (PDLSCs) as normal control, were performed. The total phenol and flavonoid contents were determined using the spectrophotometric method, and identification and quantification of polyphenols were performed using high-performance liquid chromatography (HPLC-PDA). Cytokinesis-block micronucleus assay showed that both extracts significantly increased micronucleus (MN) frequency in the PBLs treated with all tested concentrations (10, 50, 100 and 250 μg/mL), except the lowest concentration (10 μg/mL) of A. vulgaris. All the concentrations of A. alba significantly influence the nuclear division index (NDI). In treatment against MMC, the extracts dose-dependently reduced MN frequencies and NDI values in comparison with the positive control. A. alba extract exhibits significant cytotoxic activity in SW-480 cells, while A. vulgaris induces cytotoxic activity only in co-treatment with MMC after long-term exposure. Both extracts did not significantly affect the viability of the PDLSCs. The phytochemical analysis showed that the extracts contained large amounts of total phenols and flavonoids. The most abundant polyphenolic compounds were chlorogenic acid and quercetin-3-O-glucopyranoside, while 2,5-dihydroxybenzoic acid present in higher amount and detected only in A. alba extract. High concentrations of certain polyphenolic compounds detected in A. vulgaris and A. alba extracts can be a significant factor for obtaining genotoxic, cytotoxic and protective activities.

The susceptibility of Helicobacter pylori to three essential oils (EOs), 12 naturally occurring monoterpene hydrocarbons, oxygenated and phenolic monoterpenes and three reference antibiotics were studied. Classification and comparison of essential oils and monoterpenes on the basis of their chemical composition and antibacterial activity were made by the utilization of principal component analyses (PCA) and agglomerative hierarchical clustering (AHC). The most abundant compound in the Thymus glabrescens Willd. and Thymus pulegioides L. EOs is geraniol (33.8% and 52.5%), while the main constituent in Satureja kitaibelii Wierzb. ex Heuff. EO is limonene (16,1%). The compound that was the most active against H. pylori was carvacrol. EOs of T. glabrescens and S. kitaibelii exhibit higher antibacterial ability in comparison with all monoterpenes, except carvacrol, probably based on the concept of synergistic activity of essential oil components. PCA separated essential oils based on chemical composition and explain 96.5% of the total variance in the first two principal components. Essential oils, phenolic monoterpenes and two antibiotics were classified in the same sub-cluster within AHC analyses. EOs of T. glabrescens and S. kitaibelii can be used to treat infections caused by H. pylori, as a potentially effective, cheap and safe natural products. Further research of antibacterial activity of selected monoterpenes, essential oils and standard antibiotic combinations, as well as clinical study are required.

The effect of dietary genistein on performance, intestinal morphology, caecal Lactobacillus spp. count, and tibia composition in broiler chickens after 21 and 37 days of feeding was investigated. A total of 360 Cobb 500 broiler chickens (21 days old) were randomly allocated to five treatments with six replicates of 12 birds each. They were fed a basal diet (C) or a basal diet supplemented with 200 (T1), 400 (T2), 600 (T3) and 800 (T4) mg genistein/kg of feed. Genistein supplementation did not affect feed intake, but improved bodyweight, weight gain and feed conversion ratio (FCR) after 21 days, while 600 mg/kg led to a significant increase in FCR after 37 days of feeding. Plasma triglyceride level decreased with dietary genistein after 21 days, while increases were found in T3 and T4 groups after prolonged supplementation. Significantly improved duodenal and jejunal villus length and width, crypt depth and villus/crypt ratio were observed after the first and the second finishing periods, respectively, while adverse effects were found in the ileum for both periods. At 42 days old, greater spleen and heart weights were measured in broilers fed diets with 800 mg/kg than in other broiler groups. The shorter genistein supplementation period (21 days) of 200 and 400 mg/kg had a positive effect on tibia wet weight, ash and calcium (Ca) content, while 37 days of the higher genistein doses administered to the T2, T3 and T4 birds significantly increased caecal lactic acid bacteria (LAB) counts. Thus, recommended doses should not exceed 400 mg/kg. Correspondence Address: Glisic, M.; Department of Food Hygiene and Technology, Faculty of Veterinary Medicine, University of BelgradeSerbia; email: glisic.mica@gmail.com

Background: Since the beginning of the HIV/AIDS epidemic, 75 million people have been infected with the HIV and about 32 million people have died of AIDS. Investigation of the molecular mechanisms critical to the HIV replication cycle led to the identification of potential drug targets for AIDS therapy. One of the most important discoveries is HIV-1 protease, an enzyme that plays an essential role in the replication cycle of HIV. Objective: The aim of the present study is to synthesize and investigate anti-HIV-1 protease activity of some chalcone derivatives with the hope of discovering new lead structure devoid drug resistance. Methods: 20 structurally similar chalcone derivatives were synthesized and their physico-chemical characterization was performed. Binding of chalcones to HIV-1 protease was investigated by fluorimetric assay. Molecular docking studies were conducted to understand the interactions. Results: The obtained results revealed that all compounds showed anti-HIV-1 protease activity. Compound C1 showed the highest inhibitory activity with an IC50 value of 0.001 μM, which is comparable with commercial product Darunavir. Conclusion: It is difficult to provide general principles of inhibitor design. Structural properties of the compounds are not the only consideration; ease of chemical synthesis, low molecular weight, bioavailability, and stability are also of crucial importance. Compared to commercial products the main advantage of compound C1 is the ease of chemical synthesis and low molecular weight. Furthermore, compound C1 has a structure that is different to peptidomimetics, which could contribute to its stability and bioavailability.

Main goal of the most breeding programs is to develop highly adaptive hybrids in various environments, and the most important limitation are complex interactions between genotype, environment and management. Every hybrid breeding program follows certain strategy for new hybrid development. One possible strategy is to develop hybrids with lower adaptability, achieving best performance in “high input” environments (breeding for “race-horses”). However, another approach is to breed for hybrids with higher adaptability and stable performance across a wide range of environments (breeding for “work-horses”). High stability needs to be accompanied by high yield performance to insure profits, so stability should be monitored along with performance in breeding trials. Aim of this research was to analyze the new germplasm developments and their performances in the pre-registration trials in Turkey by the means of BLUP and GGE models. Heritability estimates for grain yield ranged from 0.58 to 0.85, and relative stability of all hybrids and checks is detected across all years. Cause of the high estimates of G×L interaction were crossovers of genotype performances across locations. The location Altinova was the least stable location across years. One hybrid was selected as a future check based on stability parameters across environments. As G×E interaction remains the greatest challenge in modern maize breeding, more research is needed in this field. Therefore mixed- model based approach is a valuable tool for analysis of genotype performances in maize breeding trials.

The aim of this study, besides providing new distributional data on riffle beetle Macronychus quadrituberculatus Muller, 1806 in Serbia, is a contribution to the knowledge on the species ecology. During extensive field research (2013–2016 period) a total of 415 zoobenthic samples were processed. Macronychus quadrituberculatus was found at only seven sampling sites in Serbia; all of them are situated in the Pannonian Plain or its bordering regions. Although the species moderate tolerance to organic pollution and anthropogenic pressure was noted, due to scarce findings and a low number of specimens sampled, M. quadrituberculatus regarded as rare and vulnerable species in Serbia and the Balkan Peninsula as well.

Chamomile (Matricaria chamomilla L.) is a herbaceous plant from family Astereaceae, that has a long history of use in traditional medicine. The first written report of chamomile was found in Ancient Egypt: crushed chamomile flower-heads were used to relieve skin inflammation, prevent dermatitis, and in cosmetic preparations. The subject of this study was to examine the ethnopharmacological application of chamomile in the Pirot County of Serbia. The study was conducted in the form of a population survey on the population of four municipalities: Pirot, Babusnica, Bela Palanka, and Dimitrovgrad. This study shows that chamomile has been one of the ten best known medicinal plant species in Pirot County. The most common way of the application was a herbal tea for oral use against cold, stomach diseases, throat infection, and mild sedation. Externally, in the form of compresses, it has been used against eye swelling, skin inflammation, and infections, while the oily form was used in the treatment of wounds and burns.

Second year after the fire broke out on Vidlič Mountain during the summer of 2007, the changes of burnt down beech wood vegtation were recorded. Fitocenological research of the beeech wood habitat at the location of Visoka Stena two years after the fire, was carried out in accordance with Braun-Blanquet method. The results of the terrain research were given in the form of fitocenological table. It was noticed that the most present species were Rubus idaeus, Epilobium angustifolium, Galium mollugo, Doronicum columnae, Lactuca muralis and Securigera varia. Turf grass such as Brachypodium sylvaticum, Poa nemoralis, Poa badensis, Festuca varia, Festuca valesiaca and Agrostis capillaris was also present which is a significant factor in the succession process as grass is a powerful colonist. The conclusion was that revitalization of the beech wood depends on how much the beech trees had been destroyed by the fire.

Choreutis nemorana (Hübner, 1799), the fig-tree skeletonizer moth, is a widespread species in the Mediterranean region, which has rapidly expanded its distribution area northwards and eastwards during the last 15 years. This paper reports the localities in Serbia where the C. nemorana was recorded during 2019-2020, literature data and own observations on its developmental stages, morphological characteristics, biology and symptoms of damage in the figs.

In this paper, the influence of wildfire on beech forests the third year after the fire at locality Visoka Stena on the Vidlič Mountain is followed. The results of field studies are presented in the form of phytocoenological table. The quantitative increase in the number of species diversity index in comparison to the previous two seasons was registered. Increasing number of woody and shrubby representatives of plants influences more favorable conditions for survival, germination, and further development of beech saplings in their shadows, which, with a greater wealth of nutrients in the soil than before the fire, are the good prerequisites for gradual natural regeneration of beech forests.

The paper presents phytosociological characteristics of the sessile oak and hornbeam community (Querco petraeae-Carpinetum betuli Rudski 1949. s.l.) in the area of Kosmaj. Phytosociological relevés of the investigated association were taken at altitudes ranging from 339 to 410 m, with Eastern to northeastern aspects, and the slope of 15-23°. The community occurs on deeply ilimerized soils - luvisols over flysch. The floristic composition of this phytosociological community is diverse, including plants from thermophilicmand mesothermal oak forests and some plants from beech forests. With regard to the spectrum of range types, the investigated community Querco petraeae-Carpinetum betuli Rudski 1949. s.l. has a Central European range of distribution. Regarding the bioloigical spectrum of life forms, most species are considered phanerophytes and hemicryptophytes, with an increased share of geophytes.

Xanthomonas campestris pv. campestris (Xcc), the causal agent of black rot disease of cabbage (Brassica oleracea var. capitata L.), is one of the most important bacteria which affect proper cabbage growth, leading to head weight and quality losses and thereby drastically reducing its marketing value. The pathogen is genetically diverse, which is evident from the presence of eleven races worldwide and more than thirty combinations of allelic profiles. Therefore, this study aimed to determine the allelic profiles of Serbian cabbage Xcc strains obtained in 2014. The analysis was done on three selected Xcc strains whose DNA was first amplified using polymerase chain reaction (PCR) with four housekeeping genes - P-XdnaK, fyuA, gyrB, and rpoD, then sequenced, and the obtained sequences were finally used to determine allelic profiles. Allelic profiles were determined by comparison with 33 Xcc strains obtained from different hosts and regions, whose allelic profiles had been determined previously. A non-redundant database (NRDB) from the pubMLST was used for allelic profile determination and Phyloviz software for constructing a minimum spanning tree. The obtained allelic profile of all Serbian Xcc cabbage strains was 1, 3, 1, 1 for the P-X-dnaK, fyuA, gyrB and rpoD genes, respectively. This profile is assigned as sequence type 2 (ST2) and it coincides with a Portuguese B. oleracea Xcc strain, CPBF 213, originating from B. oleracea var. costata. No connection between sequence type (ST) and the host was detected. Xanthomonas campestris pv. campestris (Xcc), prouzrokovač crne truleži kupusa (Brassica oleracea var. capitata L.) svrstava se među najvažnije fitopatogene bakterije koje utiču na pravilno razviće kupusa, dovodeći do gubitka težine glavice i njenog kvaliteta i na taj način drastično smanjuje njegovu ekonomsku vrednost. Ovaj patogen je genetički heterogen, što se ogleda kroz prisustvo dokazanih jedanaest rasa i više od trideset kombinacija alelskih profila širom sveta. Zbog svega navedenog, cilj ovog rada je determinacija alelskih profila Xcc sojeva poreklom sa kupusa prikupljenih 2014. godine. Analiza je vršena kod tri reprezentativna Xcc soja čija je DNK amplifikovana primenom lančane reakcije polimeraze (PCR) sa četiri konzervativna gena - dnaK, fyuA, gyrB i rpoD, a zatim sekvencirana i korišćena za determinaciju alelskih profila. Alelski profili su određivani poređenjem sa 33 Xcc soja izolovana sa različitih domaćina i regiona, čiji su alelski profili prethodno utvrđeni. Nonredundant baza podataka (NRDB) od pubMLST je korišćena za determinaciju alaleskih profila, a Phyloviz softver za konstrukciju Minimum Spanning stabla. Dobijeni alelski profil za sve Xcc sojeve sa kupusa iz Srbije je 1, 3, 1, 1 za gene dnaK, fyuA, gyrB i rpoD, redom. Ovaj profil je označen kao tip sekvence 2 (ST2) i podudara se sa portugalskim B. oleracea Xcc sojem CPBF 213 poreklom sa B. oleracea var. costata. Veza između tipa sekvence (ST) i biljke domaćina nije pronađena.

Lawsonia inermis is an important industrial and medicinal plant, cultivated mostly for dye production which is used in cosmetic industry. The objective of this study was establishing an efficient in vitro propagation system in order to obtain plants that will be acclimatized and grown hydroponically. The effect of different media on in vitro rooting were examined, followed by growing obtained microplants in a hydroponic culture, in a half-strength modified Hoagland nutrient solution. The highest rooting rate (79.2%) was recorded on the half-strength MS media containing 0.50 mg.L-1 IBA, with a high average number of roots (7.4). Those microplants acclimatized well in a hydroponic culture, where very rapid growth was recorded, and well developed roots and shoots were formed. After transplanting plants from hydroponic to soil, the survival rate was 100%. This is the first study reporting an acclimatization procedure in a hydroponic for the henna plant.

The high integrity of extracted DNA is necessary for the determination of human haplogroups, based on the mitochondrial DNA (mtDNA) marker sequences. The study aims were to isolate the total DNA from selected human samples and establish a protocol that gives the best yield and proper purity of DNA material for further analysis. Furthermore, human hypervariable regions - HV1 and HV2, located on mtDNA were sequenced to define human haplogroups. For extraction, samples of buccal mucosa from ten human volunteers were taken and three different protocols were used (a method with ammonium acetate, a salting-out method and GeneJET Genomic DNA Purification Kit (Thermo Fisher Scientific)). Concentration and purity of DNA were measured on BioPhotometer. Sequencing was performed by Sanger method on 3130 Genetic Analyzer (Applied Biosystems) with commercial BigDye™ Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems). Sequences were processed using Chromas and MEGA software and compared with sequence databases with BLAST tool on NCBI. Haplogroups and mutations were determined in the MITOMAP online database. Results of extraction indicated that the Purification Kit gives the highest concentration of DNA (11.86±0.63) and low purity (1.17±0.02), while a method with ammonium acetate gave the best purity (1.67±0.17). Sequencing obtained on DNA extracted with a commercial kit, gave 5 different human haplogroups U2e, T1a, J1c, H2a, K. In conclusion, it is recommended to use the Purification Kit for obtaining high integrity DNA that can be used for effective sequencing. Determination of haplogroups and haplotypes can reveal our ancestry and have important implications in molecular systematics, phylogenetics and phylogeography.

Antioxidant, antimicrobial, genoprotective, anticancer, and neuroprotective potential of acetone extract of the fruiting bodies of the edible mushroom Lactarius piperatus was studied. The antioxidant activity was evaluated using different methods (DPPH radical scavenging, superoxide anion radicals scavenging, reducing power assay, and determination of total phenolic compounds). The microdilution method was used to reveal the antimicrobial potential. The genoprotective potential was determined by Comet assay. Cytotoxic activity was tested using MTT. The capacity of the extract to inhibit acetylcholinesterase was used for determining its neuroprotective potential. The received results show that L. piperatus extract possessed potent health enhancing effects. In the antioxidant activity, IC50 was 33.97 μg/mL for DPPH radicals scavenging and 22.52 μg/mL for superoxide anion radicals scavenging, whereas the absorbance for the reducing power was from 0.0510 to 0.1451. The total content of phenolic compounds in the extract was 5.08 μg PE/mg. The testing of the antimicrobial activity showed that MIC values were from 0.039 to 10 mg/mL. For Comet assay, all concentrations of extract increased the GDI values from 0.46 ± 0.05 to 0.99 ± 0.31. L. piperatus extract expressed relatively strong cytotoxic activity with IC50 values ranging from 37.83 to 65.94 μg/mL. Finally, the percentage of inhibition of acetylcholinesterase activity of tested extract was within the range 16.75-44.35%. Our results imply that the acetone extract of L. piperatus has rather strong antioxidant, antimicrobial, genoprotective, anticancer, and neuroprotective effects; thus this mushroom represents healthy food that could be used in the pharmaceutical industry and to prevent various diseases.

Plantation cultivation of medicinal plants is expanding, and since weed control is the biggest challenge in this production, the aim of the study was to examine the possibility of their control by mulching. In addition, herbicide metamitron was included in the study, the use of which in such crops is acceptable in some countries. Angelica archangelica L. was chosen as the experimental crop as its essential oil from the root is one of the most appreciated on the market, due to which an increase in its production areas can be expected. The experiment was set up in five treatments (completely random block system with four replications), as follows: agrotextile waterproof foil, sawdust, herbicide metamitron, control without weeds (positive) and control with weeds (negative). The highest weediness was recorded in the negative control, and in the treatment with herbicide metamitron. Sawdust mulch resulted in a reduction in weediness, compared to the previous two treatments. Agrotextile foil and positive control were free of weeds. Thus, the best efficiency in weed control was achieved by applying agrotextile mulch foil, which completely prevented weed germination (efficiency 100%), a much weaker effect was achieved by using sawdust (efficiency 51.28%), while metamitron reduced fresh weed biomass by only 26.71%. The obtained results show that mulching as a physical method of weed control in angelica crop is more suitable, in comparison with the application of the herbicide metamitron, if the crop is dominated by grass species, which are not in the spectrum of action of this herbicide. The significance of these results is even greater knowing that in our country metamitron, as well as other herbicides, is not registered for use in medicinal crops.

Rosa canina, commonly known as the dog rose, is a deciduous shrub from family Rosaceae. The fruit of dog rose has a long tradition of culinary and medicinal use worldwide. The aim of this study was to analyze the ethnopharmacological application of rose in Pirot County (Southeastern Serbia). The study was conducted in the form of surveys among the population in four municipalities: Pirot, Babušnica, Bela Palanka, and Dimitrovgrad. It was noticed that rose is one of the ten best known medicinal plants in Pirot County. The fruit, named rosehip, is usually used in the folk medicine of the County in the form of herbal tea instead of coffee. Fresh rosehip contains a large amount of vitamin C and is thus usually consumed by the people of the Pirot County to prevent and treat colds, influenza, vitamin C deficiencies, and for disease prevention.

Three subterranean leptodirine leiodid taxa, viz., Bozidaria Ćurčić & Pavićević gen. nov., Bozidaria serbooccidentalis Ćurčić & Pavićević gen. et sp. nov. and Proleonhardella (Proleonhardella) tarensis Ćurčić & Pavićević sp. nov., are described and diagnosed. Bozidaria Ćurčić & Pavićević gen. nov. belongs to the phyletic series of “Leonhardella”. The new beetle taxa differ from their closest relatives in numerous morphological characters. They most likely belong to phyletic lineages of Pliocene age. The new leiodid taxa are endemic to the Dinaric mountain chain of western Serbia. Keys to the leptodirine leiodid genera of the phyletic series of “Leonhardella” and to the taxa of the genus Proleonhardella Jeannel, 1910 are included.

Stressful events experienced during early life are associated with increased vulnerability of developing psychopathology in adulthood. In the present study, we exposed 9-day-old Wistar rats to 24 h maternal deprivation (MD) with the aim to investigate the impact of early life stress (ELS) on morphological, biochemical, and functional aspects of the prefrontal cortex (PFC), a brain region particularly sensitive to stress. We found that in the superficial medial orbital cortex (MO), young adult male rats had reduced density of GAD67 and CCK immunopositive cells, while the rostral part of the ventral lateral orbital cortex (roVLO) showed a decrease in the density of GAD67 immunopositive cells in both superficial and deep layers. In addition, the superficial rostral part of area 1 of the cingulate cortex (roCg1) and deep prelimbic cortex (PrL) was also affected by MD indicated by the reduction in PV immunopositive cellular density. Furthermore, MD induced upregulation of brain-derived neurotrophic factor (BDNF), while it did not affect the overall expression of Iba1 in neonatal or young adult PFC as measured by Western blot, however, microglial activation in young adult MD rats was detected immunohistochemically in deep layers of MO and infralimbic cortex (IL). Interestingly, when young adult male rats were subjected to a behavioral flexibility test in a T-maze, MD rats showed a subtle impairment in T-maze reversal learning indicating a mildly affected PFC function. Taken together, our findings demonstrated that MD reduced the density of interneurons and induced microglial activation, in particular, PFC areas at young adulthood, and could alter synaptic plasticity accompanied by PFC dysfunction.

Plant hormones regulate numerous developmental and physiological processes. Abiotic stresses considerably affect production and distribution of phytohormones as the stress signal triggers. The homeostasis of plant hormones is controlled by their de novo synthesis and catabolism. The aim of this work was to analyse the contents of total and individual groups of endogenous cytokinins (CKs) as well as indole-3-acetic acid (IAA) in AtCKX overexpressing centaury plants grown in vitro on graded NaCl concentrations (0, 50, 100, 150, 200 mM). The levels of endogenous stress hormones including abscisic acid (ABA), salicylic acid (SA) and jasmonic acid (JA) were also detected. The elevated contents of total CKs were found in all analysed centaury shoots. Furthermore, increased amounts of all five CK groups, as well as enhanced total CKs were revealed on graded NaCl concentrations in non-transformed and AtCKX roots. All analysed AtCKX centaury lines exhibited decreased amounts of endogenous IAA in shoots and roots. Consequently, the IAA/bioactive CK forms ratios showed a significant variation in the shoots and roots of all AtCKX lines. In shoots and roots of both non-transformed and AtCKX transgenic centaury plants, salinity was associated with an increase of ABA and JA and a decrease of SA content.