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The zoonotic risk of hepatitis E virus (HEV) is well established. The HEV seroprevalence rates vary according to geographical region, assays used, and study cohorts. HEV infection is still underdiagnosed, implying the need to evaluate the disease's burden in the general population and specific risk groups, such as professionally exposed. Close contact with various animal reservoirs such as pigs, rabbits, sheep, dogs, wild boars, and deer has been associated with higher anti-HEV seroprevalence as a part of occupational exposure. While exact transmission routes remain to be determined, some general preventive measures such as proper hand hygiene, the usage of personal protective equipment, and the thermal processing of food before consumption should be followed. A “One-Health” multisectoral approach should be implemented to achieve optimal health and well-being outcomes, recognizing the interconnections between humans, animals, plants, and their shared environment, in which a vaccine against the zoonotic genotypes 3 and 4 and swine vaccination should be considered as a possible public health measure. This opinion review comprehensively addresses the HEV burden of professional exposure for butchers, slaughterhouse workers, veterinarians, farmers, hunters, and forestry workers delineates the current limits of protective work measures, and tackles future directions.

Fungi are an important source of polysaccharides (PSH) and phenolic compounds (PC). Numerous studies have highlighted the beneficial effects of fungal consumption, but the impact of submerged cultivated mycelia (M) and filtrate (F) has not been fully investigated. We aimed to investigate the cytotoxic activity of isolated crude PSH and exopolysaccharides (ePSH) of submerged cultivated M and F of edible

Multidrug resistance of human isolates of bacteria that cause vaginal infections is one of most recent topics in scientific investigations along with the search for novel antimicrobial drugs originating from nature. Monitoring of antimicrobial activity of traditionally used herbal essential oils may give us basic perspective and directions for future studies design. Antimicrobial activity of commercial essential oil (EO) of Origanum vulgare against human clinical isolates of bacteria and Candida was determined by microdilution method, obtaining minimum inhibitory concentration (MIC) and minimum bactericidal/fungicidal concentration (MBC/MFC) (CLSI protocol). Both investigated strains of Candida as well as all bacterial strains, except P. aeruginosa, showed susceptibility to investigated EO. The most susceptible strain was Gram positive S. aureus1 strain unlike S. aureus2 which indicates strain specificity. Both Gram negative E. coli isolates (E. coli1 and E. coli2) showed resistance to all tested antibiotics in this study, and susceptibility to oregano EO at 12.5 ?l/ml. GC/MS analysis identified a total of 35 components (96.13%) in commercial oregano EO, with the most dominant phenol carvacrol (64.12%). Our data suggest that oregano EO possess potent antimicrobial activity and that human vaginal isolates shows strain specificity in antimicrobial susceptibility.

BACKGROUND: High intraoperative PEEP with recruitment manoeuvres may improve perioperative outcomes. We re-examined this question by conducting a patient-level meta-analysis of three clinical trials in adult patients at increased risk for postoperative pulmonary complications who underwent non-cardiothoracic and non-neurological surgery. METHODS: The three trials enrolled patients at 128 hospitals in 24 countries from February 2011 to February 2018. All patients received volume-controlled ventilation with low tidal volume. Analyses were performed using one-stage, two-level, mixed modelling (site as a random effect; trial as a fixed effect). The primary outcome was a composite of postoperative pulmonary complications within the first week, analysed using mixed-effect logistic regression. Pre-specified subgroup analyses of nine patient characteristics and seven procedure and care-delivery characteristics were also performed. RESULTS: Complete datasets were available for 1913 participants ventilated with high PEEP and recruitment manoeuvres, compared with 1924 participants who received low PEEP. The primary outcome occurred in 562/1913 (29.4%) participants randomised to high PEEP, compared with 620/1924 (32.2%) participants randomised to low PEEP (unadjusted odds ratio [OR]=0.87; 95% confidence interval [95% CI], 0.75-1.01; P=0.06). Higher PEEP resulted in 87/1913 (4.5%) participants requiring interventions for desaturation, compared with 216/1924 (11.2%) participants randomised to low PEEP (OR=0.34; 95% CI, 0.26-0.45). Intraoperative hypotension was associated more frequently (784/1913 [41.0%]) with high PEEP, compared with low PEEP (579/1924 [30.1%]; OR=1.87; 95% CI, 1.60-2.17). CONCLUSIONS: High PEEP combined with recruitment manoeuvres during low tidal volume ventilation in patients undergoing major surgery did not reduce postoperative pulmonary complications. CLINICAL TRIAL REGISTRATION: NCT03937375 (Clinicaltrials.gov).

Ionic liquids (ILs) are well known for their physicochemical properties that recommend them for many purposes. However, many ILs are not environmentally friendly. Bearing these facts in mind, a series of imidazoliumand salicylate-based ILs with low general toxicity were designed and their pharmacological potential studied. Herein, their antiproliferative effect against human cancer cell lines and antimicrobial activity on selected Gram-positive and Gram-negative bacteria and Candida strains are presented. ILs with 1-butyl- -3-methylimidazolium or imidazolium cation (IL 1 and compound 5), with the lowest dipole moments and highest lipophilicity, exerted highest cytotoxicity against colon and/or lung cancer cells, manifesting high selectivity to the normal cells. The most non-polar IL with the 1-butyl-3-methylimidazolium cation expressed the strongest anticancer potential, but it was also toxic against normal cells, although its cytotoxicity was less than the cytotoxic effect of commercially used chemotherapeutic agents. The same compounds, IL 1 and compound 5, expressed modest effect on the bacterial strain that causes serious lung diseases and pulmonary infections (Staphylococcus aureus) or which are included in colon cancer formation (Esherichia coli and Enterococcus faecalis). Salicylate itself was toxic against lung cancer cell line A549 and affected some Candida strains. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 172012 and Grant no. 172058]

© The Royal Society of Chemistry. The fungal genus Alternaria Nees 1816 includes the most prevalent pathogenic species that can cause crop diseases such as blight, black spot, and dark leaf spot. In accordance with the aim of developing modern sustainable approaches in agriculture for the replacement of synthetic and toxic substances with environmentally friendly alternatives, the objective of this study was to examine the in vitro antifungal activities of 18 newly synthesized ionic liquids (ILs) against three Alternaria strains: A. padwickii, A. dauci and A. linicola. The antifungal activities of the ILs were estimated via a microdilution method to establish minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) values. The results confirmed that 17 of the 18 ILs showed strain specificity, including good antifungal activity toward Alternaria strains, with MIC and MFC values in the range of 0.04 to 0.43 mol dm-3. The strongest antifungal effects toward all analyzed Alternaria strains were displayed by the compounds with long alkyl chains: [omim][Cl] (MIC/MFC: 0.042 mol dm-3), [dmim][Cl] (MIC/MFC: 0.043 mol dm-3), [ddmim][Cl] (MIC/MFC: 0.053 mol dm-3), [ddTSC][Br] (MIC/MFC: 0.053 mol dm-3), and [Allyl-mim][Cl] (MIC/MFC: 0.054 mol dm-3). The introduction of oxygen as a hydroxyl group resulted in less-pronounced toxicity towards Alternaria compared to the introduction of an ether group, while the contribution of the hydroxyl group was shown to be a more determining factor than the prolongation of the side-chain, resulting in overall fungicidal activity decrease. Our results indicate the possibility that the most effective ILs ([Allyl-mim][Cl], [omim][Cl], [dmim][Cl], [ddmim][Cl], [bTSC][Br], [hTSC][Br], [oTSC][Br], [dTSC][Br], and [ddTSC][Br]) could be applied to the control of plant diseases caused by Alternaria species, based on their potential as an environmentally friendly crop protection approach. Since salts based on TSC cations are significantly cheaper to synthesize, stable under mild conditions, and environmentally friendly after degradation, thiosemicarbazidium-based ILs can be a suitable replacement for commercially available imidazolium ILs.

This journal is © The Royal Society of Chemistry. This study was designed to define total protein, phenol and flavonoid content as well as LC-MS/MS phenolic profile related to antioxidant and antidiabetic activity of ethanolic (EtOH) and water extracts of G. pfeifferi and G. resinaceum. G. resinaceum water extract possessed the highest ability to scavenge DPPH and O2-, while the EtOH extract of the same species showed better activity on NO related to other extracts. The highest level of bioactive compounds was determined generally in EtOH extracts. Antidiabetic action was evaluated by the oral glucose tolerance test (OGTT) and histological examination of pancreas and liver in normoglycemic and alloxan-induced diabetic animals. Histological examination of pancreatic tissue demonstrated that G. pfeifferi extracts have protective effects. To conclude, analysed extracts could be considered as a promising candidate for further research with the aim to promote antidiabetic activity, which is for the first time reported for G. pfeifferi.

Kirurško liječenje deformiteta šaka bolesnika s buloznom epidermolizom, unatoč stalnomu napretku znanstvenih spoznaja, još je i danas obilježeno brojnim prijeporima. Cilj istraživanja je na temelju medicinske dokumentacije utvrditi karakteristike, specifičnosti i ishode svih bolesnika s buloznom epidermolizom koji su kirurški liječeni zbog deformiteta šake u Zagrebu u razdoblju od 1994. do 2009. godine. Zagrebački kirurški protokol rekonstrukcije (ZSRP) deformiteta šake u bolesnika s buloznom epidermolizom (EB) jedinstven je alat standardiziranih postupaka i jasno definiranog algoritma. Analizirani su značajke i ishodi u bolesnika s buloznom epidermolizom skupine bolesnika liječenih u razdoblju od 1994. do 2001. sa skupinom bolesnika koji su kirurški liječeni od 2002. do 2009. godine (s follow upom od 2 godine poslije termina operacije) s obzirom na to da su od 2002. godine u kiruršku praksu uvedene nove smjernice s brojnim modifikacijama postupka (ZSRP). Primjena Zagrebačkog kirurškog protokola rekonstrukcije deformiteta šake u bolesnika s buloznom epidermolizom sa svojim inovativnim sastavnicama ima praktične prednosti u usporedbi s prethodno primjenjivanim kirurškim postupcima zbog svoje veće učinkovitosti uz redukciju ukupnih troškova liječenja. Uzimanjem presatka kože po metodi PRST postiže se brže cijeljenje donorskog područja, uz smanjenje rizika očekivanih komplikacija u usporedbi s klasičnim tehnikama uzimanja STSG presatka kože dermatomom ili elektrodermatomom. Prema našem dosadašnjem 20-godišnjem iskustvu primjerena kirurška korekcija deformiteta šaka u bolesnika s EB-om omogućava uspješno višekratno ponavljanje kirurškog postupka koji je indiciran kao posljedica očekivane progresije lokalnog statusa zbog prirode same bolesti., Surgical treatment of the deformity in a handful of patients with epidermolysis bullosa (EB) is still marked by numerous controversies today despite constant advancement of scientific knowledge. The aim of the study was to determine, on the basis of medical documentation, the characteristics, specificities and outcomes of all EB patients surgically treated in Zagreb from 1994 to 2009. The Zagreb Surgical Reconstruction Protocol (ZSRP) of hand deformity in EB patients is a unique tool of standardized procedures and a clearly defined algorithm. Characteristics and outcomes were compared between EB patients treated from 1994 to 2001 and EB patients treated surgically from 2002 to 2009 (with 2-year follow-up after surgery), given that new guidelines with numerous modifications to the procedure (ZSRP) had been introduced into surgical practice from 2002. Using ZSRP for reconstruction of hand deformity in EB patients with its innovative components was found to have practical advantages compared to the previously applied surgical procedures because of its higher efficiency while reducing the overall cost of treatment. This method resulted in faster healing of the donor area, reducing the risk of expected complications compared to the conventional split-thickness skin grafting techniques with a dermatome or electrodermatome. According to our 20-year experience, appropriate surgical correction of hand deformity in EB patients enables successful repeated surgery, which is indicated as a result of the expected progression of local status due to the nature of the disease itself.

This study was focused on in vitro screening of the total antioxidant activity of the selected extracts of the mushroom Coprinus comatus and quinic acid, one of their antidiabetic ingredients, by an uncommon electrochemical assay. Indeed, direct current (DC) polarographic HydroxoPerhydroxo Mercury(II) Complex (HPMC) assay based on decrease of anodic limiting current originating from HPMC formation in alkaline solutions of hydrogen peroxide at potential of mercury dissolution, observed upon gradual addition of antioxidants, was applied herein for the estimation of the natural products' antioxidativity. Quinic acid was found to exhibit most promising antioxidant potential (4.0 ± 0.2%µL-1) being ≈ 2-fold more active than the screened C. comatus extract samples. Actually, such a finding puts some light on the antioxidativity of cyclic polyols, well understimated class of organic compounds, compared to aromatic (poly)phenolics. As a low cost, easy-to handle and accurate this polarographic assay may be thoroughly recommended for much broader use. [Formula: see text].

Many lignicolous mushroom species are used as a food supplement and may represent an alternative treatment of Alzheimer's disease (AD). This study aimed to evaluate acetylcholinesterase inhibition (AChEI) of Stereum hirsutum together with antioxidant activity (AO) and cytotoxic activity against HepG2 cells. Different extracts (water, ethanol, methanol, polysaccharide) were analyzed, with respect to their mineral composition and chemical content. Ethanol extract was the most potent in AChEI (98.44 %) and demonstrated cytotoxic activity (91.96 % at 900.00 μg/mL), while the highest AO was demonstrated for polar extracts (methanol and water) as well. These activities may be attributed to determined phenolics (hydroxybenzoic and quinic acid) and fatty acids (FA), while biflavonoid amentoflavone may be responsible for cytotoxic activity. The most prevalent FA was linoleic (40.00 %) and the domination of unsaturated FA (UFA) (71.91 %) over saturated (26.96 %) was observed. This is the first report of AChEI of S. hirsutum extracts and first detection of amentoflavone. Due to high amount of UFA and well-expressed AChEI, this species can be considered as a potent food supplement in the palliative therapy of AD.

Fungi could absorb heavy metals, metalloids, or radionuclides, thus fungal species possess great potential in bioremediation. Since fungi absorb the vanadium, in the present study ability of Coprinellus truncorum mycelia for vanadate uptake and its intracellular metabolism were investigated. The submerged cultivated C. truncorum was exposed to a rising concentration of vanadate. 31P NMR spectroscopy was used to investigate phosphate metabolism of the mycelium, while the status of vanadium in the cell was followed by 51V NMR spectroscopy. The mycelium could grow, and overcome vanadate presence, up to the concentration of 1.6 mM in the submerged medium. 31P NMR measurements pointed out that vanadate induced changes in the concentration of the crucial metabolite containing phosphorus, particularly sugar phosphates. The major result of vanadate action is evinced through an appearance of a signal positioned at around 2.8 ppm, and an increased signal of hexose- phosphates. Using 51V NMR spectroscopy the presence of vanadate monomer in the mycelia of the fungal cell was confirmed.

© The Royal Society of Chemistry. In this work, new protic ionic liquids (PILs) with 1-ammonium-2-propanol cation and nine different anions: formate (For), acetate (Ac), lactate (Lac), trifluoroacetate (TFA), chloroacetate (ClA), trichloroacetate (TClA), 3-chloropropionate (3-ClP), 4-chlorobutyrate (4-ClB) and mandelate (Man) were prepared in order to study their antimicrobial activity and possible application for fungi and bacteria removal from deteriorated paper heritage. Ten filamentous fungal strains isolated from specific pigmented area of the damaged books: Trichoderma sp., Cladosporium sp., Penicillium sp.(1-3), Penicillium citrinum, Aspergillus sp.(1,2), Aspergillus flavus, Fusarium graminearum, eight Gram positive and Gram negative ATCC bacterial strains: B. subtilis(6633), S. aureus(6538), E. faecalis(19433), K. rhizophila(9341), E. coli(11229), S. enteritidis(13076), P. mirabilis(12453), P. aeruginosa(15692) and two yeast Candida strains: Candida albicans(ATCC 10231) and Candida albicans(L) were used in this study. The results indicated that antimicrobial activity of selected ionic liquids is significantly affected by the size and specific functional groups in the anion structure. These facts opened the possibility for molecular design of new ionic liquids with strong inhibition properties against the specific bacterial, mould and yeast strains. The significant antimicrobial properties observed in this research suggest that studied PILs may have potential applications in the paper art and artefact cleaning and conservation replacing thus, conventional solvents and organic substances that are toxic for humans and environment.

Saprotrophic basidiomycetes play a crucial role in leaf-litter decomposition, especially in nitrogen-limited boreal and temperate forests. Populations of this group of fungi have been inadequately investigated. We examined the populations of three different saprotrophic species ( Mycetinis alliaceus , Marasmius rotula and Gymnopus androsaceus ) in forests in Serbia and Montenegro. To determine the distribution of genets at each of the three investigated sites, molecular analysis was conducted using the inter-simple sequence repeats (ISSR) method. Seven to fifteen genets (genotypes, individuals) were identified on each site and the majority of them were represented by a single sporocarp. The sizes of the genets with two or more sporocarps were estimated to range from 0.3 to 4.0 m. Results obtained in this study suggest that populations of these three species can consist of numerous and relatively small genets. https://doi.org/10.2298/ABS190218025B Received: February 18, 2019 ; Revised: April 4, 2019; Accepted: April 9, 2019; Published online: April 10, 2019 How to cite this article: Boskovic EV, Galovic VO, Karaman MA. Spatial distribution of genets in populations of Saprotrophic basidiomycetes, Mycetinis alliaceus , Marasmius rotula and Gymnopus androsaceus , from Serbian and Montenegrin forests. Arch Biol Sci. 2019;71(3):435-41.

© 2018, Lithuanian Research Centre for Agriculture and Forestry; Vytautas Magnus University. All rights reserved. Clove rot caused by Fusarium spp. is a very important disease of common garlic (Allium sativum L.) occuring in many areas of the world. However, there is a lack of data about biocontrol of these pathogens. Bacillus species are attractive for research due to their potential use in the biological control of fungal diseases. The aim of this study was to select effective biocontrol agents from a series of indigenous Bacillus spp. isolated from soil. Bacterial isolates positive for hydrolytic enzymes production were screened for antifungal activity against Fusarium spp. isolated from infected garlic cloves. Polymerase chain reaction (PCR) analyses were used for molecular identification of bacterial (16S rDNA gene) and fungal (EF-1α gene) isolates, and detection of biosynthetic genes for antimicrobial lipopeptides (surfactin, iturin, bacillomycin D and fengycin) in Bacillus spp. The obtained results confirmed the presence of Fusarium tricinctum, F. oxysporum f. sp. cepae, F. proliferatum, F. acuminatum and F. verticillioides as the causal agents of garlic clove rot. Four bacterial isolates identified as Bacillus subtilis exhibited the highest antagonistic effect during in vitro testing of antifungal activity (up to 71% reduction in fungal growth), and caused a significant suppression of garlic clove infection (up to 58% reduction in rot symptoms) in situ. Analysis of the antifungal compounds involved in the antagonistic activity of the examined isolates revealed their ability to produce the antibiotic lipopeptide surfactin. The most effective isolates of B. subtilis could be used as potential biocontrol agents of garlic clove rot.

© 2018, University of Tehran. Salmonellae are important bacterial pathogens dispersed in the environment through wastewater and agricultural application of organic fertilizers, while infections are mainly food borne. Although routine monitoring in human, meat production and processing areas and in animal meat is conducted, little is known about Salmonella persistence in environmental samples, even though they are highly important ecological reservoirs for the spreading of pathogen and antibiotic resistance determinants. Here, we present a large-scale survey of Salmonella isolates from a variety of soils and sediments in Serbia. Among 1062 analyzed samples, 67 (6.31%) tested salmonellae positive. A number of isolates persisted in soil and sediment stored at low temperatures for up to 415 days. A third of the tested isolates exhibited atypical biopatterns unusual for salmonellae. Majority of strains (87.5%) were either sensitive or intermediate to all the 20 tested antimicrobials, while only two strains were multidrug resistant (6.25%). Notably, all isolates were sensitive to medically important antibiotics such as fluoroquinolone, ciprofloxacin and beta-lactams. Serotyping identified three subspecies, namely enterica, salamae and diarizonae and 15 serovars, with the most prevalent ones being Brandenburg (n = 9), Enteritidis (n = 8) and Wien (n = 7). With the exception of S. Enteritidis, other detected serovars are rarely present in human and animal samples. To conclude, Salmonella strains isolated from soil and sediment samples across Serbia exhibited unusual biochemical properties, low level of antibiotic resistance and high serovar diversity. Results indicate serovars uncommon in human and animal meat samples are more persistent in the outside non-host environments.

© 2018, © 2018 Informa UK Limited, trading as Taylor & Francis Group. This study outlines antioxidant and anti-AChE activities of the polysaccharide (PSH) extract from the mushroom species Amanita strobiliformis. Both the presence of α and ß glucans within the aforementioned extract was recorded. PSH extract displayed a profound scavenging activity of OH radicals (IC50 value, 11.86 ± 0.59 μg/mL) and high potential for reduction of Fe3+ ions (174.11 ± 8.70 mg eq. AA/g d.w.) being almost 48- and 5-fold more effective than mannitol and butylated hydroxytoluene used as a positive control, respectively. Compared with galanthamine (0.001 μg), the same extract exhibited a moderate anti-AChE activity (10 μg) in solid. Since purified PSH extract exhibited higher bioactivity (IC50 value 7.27 ± 0.31 μg/mL, 197.68 ± 9.47 mg eq. AA/g d.w. and 0.1 μg, respectively), it can be predominantly ascribed to the polysaccharide compounds. A. strobiliformis PSH extract may be considered as a promising resource of potent bioactive polysaccharides of natural origin successfully addressing both oxidative stress and lack of acetylcholine.

The aim of this work was to investigate nutritional profile in relation to proteins, amino acids, fatty acids and mineral composition, as well as phenolic profile of small edible fungal species Coprinellus disseminatus originated from Serbia. Total protein content in the analyzed fungal species was 9.72%. Fifteen protein fractions obtained by electrophoresis were identified in a range from 1.6 to 63.6 kDa. Chip-based separations showed the presence of protein fraction with molecular weight of 27.5 kDa that could possess antifungal activity. The total essential and non-essential amino acid contents were 29.57 and 96.69 mg/g DW, respectively. Among the essential amino acids, leucine was the most abundant. Fatty acid composition of C. disseminatus showed that polyunsaturated fatty acids (PUFA, 59.1% of total FA) predominated over saturated fatty acids (SFA, 23.1% of total FA) and monounsaturated fatty acids (MUFA, 17.9% of total FA). The dominant fatty acids were linoleic acid (56.6%), palmitic acid (13.9%), and oleic acid (12.0%). The most abundant macroelement in C. disseminatus was potassium, followed by calcium and magnesium, while iron dominated in microelements. Eight phenolic compounds were quantified in methanolic extract of C. disseminatus by LC-MS/MS with the highest amount of p-hydroxybenzoic acid and p-coumaric acid reaching 9.46 ± 0.2 µg/g DW and 7.8 ± 0.1 µg/g DW, respectively.

Aims This study aimed to investigate the application of Origanum vulgare L. essential oil (EO) as an alternative antimicrobial agent against vaginal infections. Methods and Results Microdilution methods were applied for the detection of the minimal inhibitory (MIC) and the minimal bactericidal/fungicidal (MBC/MFC) concentration of 15 clinical strains originating from women with symptoms of vaginal infection. Optical density determination (OD) was used for detection of the Candida growth rate. Staining with DAPI was used to detect the influence of EO on nuclear condensation and fragmentation, while the brine shrimp bioassay was used to determine the toxicity of EO. Chemical composition analysis was done using GS-MS. According to the MIC and MBC/MFC values, the most susceptible strains to EO were: Escherichia coli 1, E. coli 2, Staphylococcus aureus 3 and Candida albicans 1–3. Inhibition of C. albicans filamentation was detected at 0·45 μl ml−1. Conclusions The obtained inhibition (%) from Candida growth curves points to a shorter period of time (24 h) for determining IC50 as MIC and IC99 as MFC value. These values could be recommended as valid parameters for the faster detection of the effectiveness of EO on Candida isolates. Significance and Impact of the Study Examination of potential of the O. vulgare EO as a main antimicrobial constituent within vaginalettes in gynaecological practice.

Matthiola longipetala (Vent) DC, commonly known as “night-scented stock” or “evening stock”, is the most widely cultivated species of the genus Matthiola in the family Brassicaceae. It is a common garden flower, available in a variety of colors, many of which are heavily scented and also used in floristry. An elevated incidence of Fusarium was observed during a routine quality control seed assay of M. longipetala obtained from a private production facility in Đurđevo (South Backa District) in 2018. Fungal infection was noticed on an average of 30% of the tested seed, followed by a reduction in germination. The infected seed was covered with white to beige mycelium. Prior to isolation, seeds were surface sterilized for 10 min with a 1% sodium hypochlorite solution to reduce contaminants, washed twice in sterile water, and plated on potato dextrose agar (PDA). After 7 days of incubation at 25°C under a 12-h/12-h photoperiod of fluorescent light, 14 Fusarium (JBL4089/1 to 14) isolates were single spored and subcultured on both PDA and carnation leaf agar (CLA). Pathogenicity was performed in vitro using a modified PDA slant method in a test tube (Porter et al. 2015). A piece of mycelium of each isolate grown on PDA for 7 days was placed at the bottom of each tube, and dried M. longipetala seed was carefully placed 2 cm above the inoculum. After 10 days, fungal mycelia of 14 isolates completely covered the seedlings, causing seed rot and seedling decay. The Fusarium was reisolated on PDA and used for further analysis in order to morphologically identify the species. Isolate JBL4089/2 formed abundant, loosely floccose, whitish aerial mycelium with beige pigmentation. After transfer to CLA, the isolate formed macroconidia with a tapered and elongated apical cell and prominent foot-shaped basal cell, which were typically four to five septate, with average dimensions of 21 to 60 × 2.8 to 4.6 µm. The isolate formed chlamydospores, but microconidia were not observed. Based on the morphological characteristics, isolate JBL4089/2 was identified as Fusarium equiseti (Corda) Sacc. according to Leslie and Summerell (2006) and Gerlach and Nirenberg (1982). Identification of isolate JBL4089/2 was confirmed by amplification and sequencing of a portion of the translation elongation factor-1 alpha (EF-1α) gene. Total DNA was extracted directly from fungal mycelium with a DNeasy Plant Mini Kit (Qiagen, Hilden, Germany), and PCR amplification was performed with primer pair EF-1/EF-2 (O’Donnell et al. 1998). Sequence analysis of the EF-1α gene revealed 100% nucleotide identity of isolate JBL4089/2 (GenBank accession no. MK061538) with the EF-1α sequences of two F. equiseti isolates from Canada (KU587617 from Pisum sativum and MH315936 from Glycine max) and a Hyssopus officinalis isolate (MK061540) of F. equiseti from Serbia. To our knowledge, this is the first report of F. equiseti as a causal agent of seed rot on M. longipetala in Serbia. The presence of the pathogen could cause significant economic losses in M. longipetala production, and for that reason control strategies for the management of the disease should be implemented.

© 2020 Wiley-VHCA AG, Zurich, Switzerland The aim of this research work was to study the bioactivity potentials (antioxidant, anti-inflammatory, and enzyme inhibitory) of ethanol (EtOH), water (H2O) and chloroform (CHCl3) extracts of G. applanatum, G. lucidum, G. pfeifferi and G. resinaceum as well as their mycochemical profile: the total content of phenolics (TP) and sugars (TS) and LC/MS/MS detection of phenolics. LC/MS/MS profile showed that p-hydroxybenzoic and protocatechuic acids were mostly found. The highest ABTS and DPPH activities were detected in polar G. applanatum extracts (159.84±0.59 mg TE/g d.w., IC50=0.85±0.30 μg/mL, respectively), while G. resinaceum CHCl3 extract was the most potent in NO assay (IC50=41.21±0.18−81.89±0.81 μg/mL). The highest TP and TS were generally determined in G. applanatum EtOH extracts. Enzyme inhibitory effects were determined in H2O extracts. Generally, CHCl3 extracts showed the most powerful anti-inflammatory potential. These results suggest that analyzed species are a promising source of bioactive compounds and may be considered as candidates for new food supplements or drug formulations.

This study was designed to define total protein, phenol and flavonoid content as well as LC-MS/MS phenolic profile related to antioxidant and antidiabetic activity of ethanolic (EtOH) and water extracts of

This study was focused on

Ganoderma lucidum, used in a traditional Chinese medicine, represents one of the most important medicinal mushrooms in the world, whose fruiting bodies and spores have been traditionally used because of a wide spectrum of biological activities such as antidiabetic, antioxidative, antiproliferative, cardioprotective, etc. Its ethnomedicinal importance in some parts of the Balkan region (Serbia and Croatia) is almost totally unknown and there should be more scientific investigations carried out. The aim of this work was to make a comparative study of antioxidative activities and total phenolic content of ethanol and hot water extracts of G. lucidum, collected from forests in Serbia (Morovićke šume, Fruška Gora) and Croatia (Donji Lapac, Plješevica). The present study was carried out to evaluate antioxidant potential of examined extracts via scavenging potential on ABTS, DPPH, OH∙ and NO∙ radicals, as well as of chelating effects via FRAP assay, together with determination of their total phenolic content. Results showed that both GLS extracts possessed better antiradical activities (IC50=0.23±0.01 for H2O and 2.75±0.01 μg/mL for EtOH for OH∙ and DPPH assay, respectively) than in the ABTS assay (151.40±1.07 mg TE/g d.w. for EtOH), while the phenolic content was generally equal in extracts of Serbian and Croatian samples (60.74±0.57 mg GAE/g d.w. for EtOH and 77.10±0.27 mg GAE/g d.w. for H2O, respectively). Therefore, these extracts could be considered as a good source of natural antioxidants. These results showed that examined G. lucidum extracts (especially Н2О) contain high amount of phenolic content which could significantly enhance the antiradical potential and reduce potential on iron ions. This is the first study reporting the comparison of antioxidant activities and phenolic contents of two different extracts between two G. lucidum strains from two different geographical origins from the Balkan region. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 43002]

Biocontrol using plant growth-promoting rhizobacteria (PGPR) represents an alternative approach to disease management, since PGPR are known to promote growth and reduce diseases in various crops. Among the different PGPR, members of the genus Bacillus are prefered for most biotechnological uses due to their capability to form extremely resistant spores and produce a wide variety of metabolites with antimicrobial activity. The objective of this research was to identify antagonistic bacteria for management of the plant diseases. Eleven isolates of Bacillus spp. were obtained from the soil samples collected from different localities in the Province of Vojvodina. The antifungal activity of bacterial isolates against five fungal species was examined using a dual plate assay. Bacillus isolates exhibited the highest antifungal activity against Fusarium proliferatum, Fusarium oxysporum f. sp. cepae and Alternaria padwickii, while they had the least antagonistic effect on Fusarium verticillioides and Fusarium graminearum. Molecular identification showed that effective bacterial isolates were identified as Bacillus safensis (B2), Bacillus pumilus (B3, B11), Bacillus subtilis (B5, B7) and Bacillus megaterium (B8, B9). The highest antagonistic activity was exhibited by isolates B5 (from 39% to 62% reduction in fungal growth) and B7 (from 40% to 71% reduction in fungal growth). These isolates of B. subtilis could be used as potential biocontrol agents of plant diseases. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR-31073]

This study was conducted to determine the size and spatial distribution of mycelial individuals of Marasmius rotula at one locality on Mt. Stara planina in the Republic of Serbia. Total of 12 sporocarps were collected from investigated locality (Vidlič, Stara planina). Sporocarps were distributed in four groups and distances between them were approximately 10-30 meters. Genomic DNA was extracted from each sporocarp and used for inter-simple sequence repeat (ISSR) polymorphism analysis using (GTG)5 and (GACA)4 primers. Both primers showed reproducible band patterns on agarose gels and sporocarps with identical band patterns were considered to belong to the same individual (genet) and were grouped accordingly. Grouping with both primers determined that 12 analyzed sporocarps belong to 4 distinct genets (A, B, C, D). Approximate genet diameters were 2 m for two genets (A, B) and 15 m for one genet (C) while diameter of one genet (D) was not possible to determine since it was represented only by one sporocarp. The results presented here are the first data about size and spatial distribution of genets of M. rotula. To determine whether the obtained genet sizes are general trait of an analyzed species or a special trait appeared as an effect of environmental conditions, more information on the genet distribution of other M. rotula populations is needed. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. III43002 and Grant no. III43007]

peer-reviewed, The aim of this work was to study the bioactivity of crude aqueous and ethanolic extracts of Boletus edulis prepared from caps and stipes of wild-growing basidiocarps collected from the Prijepolje region (western Serbia). The bioactivity screening included antioxidant (2,2-diphenyl-l-picrylhydrazyl [DPPH], nitric oxide, super-oxide anion*, and hydroxyl radicals and ferric-reducing antioxidant power) and antiproliferative MTT assays (human breast MCF-7 cancer cell line). In addition, all extracts were primarily characterized by ultraviolet/visible spectrophotometry to determine total phenolic and flavonoid contents. The highest anti-DPPH and anti-hydroxyl radical activity were observed in aqueous B. edulis extract from the caps (half maximal inhibitory concentration [IC50] = 50.97 μg/ mL and 2.05 μg/mL, respectively), whereas the highest anti-nitric oxide radical activity was observed in aqueous B. edulis extract from the stipes (IC50 = 10.74 μg/mL). The ethanolic extract obtained from the mushroom stipe showed higher anti-superoxide anion radical activity (IC50 = 9.84 μg/mL) and ferric-reducing antioxidant power (22.14 mg ascorbic acid equivalents/g dry weight) compared with aqueous extracts. Total phenolic content for all extracts was similar but total flavonoid content was significantly higher in the aqueous B. edulis extract from the caps (4.5 mg quercetin equivalents/g dry weight). All crude extracts showed activity against the MCF-7 cell line, with the ethanolic extract of B. edulis prepared from stipes (IC50 = 56 μg/mL) being the most potent. This is, to our knowledge, the first report of the antiproliferative effects of crude aqueous and ethanolic extracts prepared from caps and stipes of wild-growing basidiocarps of B. edulis on the human breast MCF-7 cancer cell line., This study was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia (projects III 46001 and OI 172058)

peer-reviewed, The aim of this work was to analyse mineral composition and chemical profile of two nonedible fungal species: Ganoderma lucidum and Ganoderma applanatum (Fruška Gora, Serbia) vs. their antioxidant (ABTS and A.E.A.C. assay) and cytotoxic biopotentials (MTT assay on MCF-7). Both species were analysed for their content of macro- and microelements by atomic absorption spectrophotometry, while phenolic profile of EtOH and H2O extracts was examined by LC-MS/MS technique. Both species mostly contained the following ions: K+ > Ca2+ > Mg2+ > Mn2+ > Zn2+ > Cu2+ > Cr3+. Among nine phenolic compounds, the highest content of vanillic acid was detected in G. applanatum extracts while protocatechuic acid in EtOH extract and quinic acid in H2O extract were mostly contained in G. lucidum. Ganoderma applanatum EtOH extract showed the best antioxidant activities related to its phenolic and flavonoid content. Further, the best cytotoxic effect after 72 h was observed in this extract as well., This research work was supported by Project of Ministry of Education and Sciences Republic of Serbia (Grant No.172058) as well as to the Provincial Secretariat for Science and Technological Development, Vojvodina, Serbia in the frame of project APV 114-451-849/2015-03.

Esophageal stricture is one of the most common gastrointestinal complication in epidermolysis bullosa patients. We describe the technique of esophageal stricture dilatation, with a focus on safety and procedural modifications. Case series of 6 patients diagnosed with epidermolysis bullosa who underwent esophageal dilatation in our unit. Type of epidermolysis bullosa, patient’s age, dilatation technique, dysphagia free interval and complications were recorded. Four females and two males underwent a total of 31 dilatations during a 4-year follow-up. Most common dilatation technique was “through-the-scope” with three-stage baloons without radiology control, and in one case Savary bougi. Dilatation is usually done as an outpatient procedure with patients being discharged 3–4 h after the procedure. Mean dysphagia free interval is 7 months, longer is older patients. “Through-the-scope” baloons could be the best way to perform esophageal dilatation in epidermolysis bullosa patients. The procedure is short, safe with high success rate and dysphagia free interval and minimal complications.

Tapentadol je novi lijek registriran u retard obliku za umjerenu i jaku kroničnu bol koji svojim spektrom djelovanja pokriva i nociceptivnu i neuropatsku komponentu boli. Analgetski se učinak postiže djelovanjem dvaju sinergističkih mehanizama: agonističkim učinkom na mi opioidne receptore (MOR-agonizam) i inhibicijom ponovnog preuzimanja noradrenalina (NRI). Cilj: Usporediti naša iskustva o učinkovitosti i podnošljivosti tapentadola s iskustvima drugih istraživača u svijetu. Metode: U istraživanje su uključena 92 bolesnika iz naše ambulante za bol, kojima je bilo indicirano propisati tapentadol-retard u dozi od dva puta 50 mg ili višoj. Pratili smo povoljni učinak na smanjenje bolova prema vizualno-analognoj ljestvici (VAS) te vrstu i učestalost neželjenih učinaka lijeka. Bilježene su i daljnje preporuke za prilagodbu doze odnosno prekid terapije. Rezultati: Pojava povoljnog djelovanja tapentadola na smanjenje kronične boli kod različitih bolnih sindroma pokazala se u 66 %, ispitanika. Postotak nuspojava, od blažih do težih, iskazalo je 43 % bolesnika. Pedeset i četiri % korisnika tapentadola odustalo je od terapije, 37 % zbog nuspojava, 15 % zbog nezadovoljstva kontrolom boli i 2 % zbog neodobravanja obiteljskog liječnika. Zaključak: Našim istraživanjem potvrđena je dobra učinkovitost tapentadola na smanjenje boli. Nuspojave se u ukupnom broju javljaju sličnom učestalošću kao u podatcima iz literature, i to rjeđe nego kod terapije klasičnim opioidima. Uočena je visoka stopa prekida terapije što je u prvom redu pripisano pojavi neželjenih učinaka., Tapentadol is a novel analgesic registered in slow-release form for medium-intensity and intense chronic pain, which targets both the nociceptive and neuropathic component of the pain signal. It utilizes two synergistic mechanisms: MOR – mu opioid receptor agonism and NRI – norepinephrine reuptake inhibition. The goal of this study was to compare the effectiveness of pain management and tolerability of tapentadol in our clinical experience with existing reports from abroad. The study included 92 patients from our pain management clinic who were prescribed slow-release tapentadol in doses of at least 50 mg twice daily. We tracked the effectiveness of pain management recorded using the visual analog scale, as well as the appearance and type of adverse effects. Further directions for dose adjustment or discontinuation of therapy were also recorded. In our experience, 66% of patients reported a satisfactory analgesic effect. The adverse effect rate was 43%; 54% of patients discontinued tapentadol therapy, i.e. 37% due to adverse effects, 15% because of inadequate pain management, and 2% because of a negative attitude of their primary care physician. Our study confi rmed the effi cacy of pain management with tapentadol. The total adverse effect rate was comparable to reports found in the literature and was lower than with classical opioid therapy. A relatively high therapy discontinuation rate was noted primarily due to adverse effects.