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Pears are one of the oldest and the third most important fruit species grown in temperate regions. They are consumed because of their nutritional and health benefits, in fresh form or as various processed products. This article resolves the etiology of the Penicillium-like mold symptoms on pear fruits in Serbia. Samples of pear fruits with blue mold and other Penicillium-like mold symptoms were collected in Serbia from 2016 to 2019, from four storages. The recovered isolates were identified and characterized according to a polyphasic approach. Morphological and physiological analyses were performed on three media and five temperatures, respectively. Four loci (internal transcribed spacer, beta-tubulin, calmodulin, and DNA-dependent RNA polymerase II second largest subunit) were used for sequencing, genetic identification, and phylogenetic analyses. The results of the identification by conventional and molecular methods were in agreement, and they revealed that the obtained isolates belong to five species: Penicillium crustosum, P. expansum, P. italicum, Talaromyces minioluteus, and T. rugulosus. In a pathogenicity test, P. crustosum, P. expansum, T. minioluteus, and T. rugulosus produced decay on artificially inoculated pear fruits, and P. italicum induced tissue response lesions. The results of this study are the first reports of T. minioluteus and T. rugulosus as postharvest pear pathogens. Also, these are the first world records of T. minioluteus, T. rugulosus, and P. italicum on fruits of European pear. Furthermore, this is the first finding of P. crustosum, P. expansum, P. italicum, T. minioluteus, and T. rugulosus on pear fruit in Serbia.

In the last several decades, amphibian populations have been declining worldwide. Many factors have been linked to global amphibian decline, including habitat destruction, pollution, introduced species, global environmental changes, and emerging infectious diseases. Recent studies of amphibian skin infections were mainly focused on the presence of chytridiomycosis, neglecting other members of the frogs' skin communities. The diversity pattern of fungal dwellers on the skin of green frogs (Pelophylax esculentus complex) was investigated. A total of 100 adults were sampled from three localities in South Banat (northern Serbia) over three consecutive years and detected fungal dwellers were identified using light microscopy and ITS and BenA gene sequencing. Structures belonging to fungi and fungus-like organisms including a variety of spores and different mycelia types were documented in the biofilm formed on amphibian skin, and are classified into 10 groups. In total, 42 fungal isolates were identified to species, section, or genus level. The difference in mycobiota composition between sampling points (localities and green frog taxa) was documented. The highest number of fungal structures and isolates was recorded on the hybrid taxon P. esculentus and locality Stevanove ravnice. Parental species showed a markedly lower diversity than the hybrid taxon and were more similar in diversity patterns and were placed in the same homogenous group. The locality Stevanove ravnice exhibited more pronounced differences in diversity pattern than the other two localities and was placed in a distinct and separate homogenous group. Among the fungal isolates, the highest isolation frequency was documented for Alternaria alternata, Aspergillus sp. sect. Nigri, Epicoccum nigrum, Fusarium proliferatum, and Trichoderma atroviride. Among the documented species, dematiaceous fungi, causative agents of chromomycosis in amphibians, were also recorded in this research with high isolation frequency. Also, some rare fungal species such as Quambalaria cyanescens and Pseudoteniolina globosa are documented for the first time in this research as microbial inhabitants of amphibian skin.

Grifola frondosa HAI 1232 was tested for ligninolytic enzyme activities and for lignin, cellulose and hemicellulose degradation during cultivation on eight common agro-forestry residues in Serbia. Wheat straw was favorable lignocellulosic for the production of Mn-dependent and Mn-independent peroxidases (2513.89 and 354.17 U L-1, respectively), while selected residues inhibited the synthesis of laccases. The highest lignin removal was observed during fermentation of blackberry sawdust (36.75%), while the highest selectivity index was recorded on oak sawdust (4.34). The dry matter loss varied between 8.17% in corn stalks and 14.16% in apple sawdust. According to the presented results, it can be concluded that G. frondosa HAI 1232 could be an important participant in various biotechnological processesdue to its high capacity to selectively degrade different agro-forestry residues.

Leaf spot diseases caused by Alternaria species have been reported worldwide in plants in the Brassicaceae family. However, there is little information on Alternaria species causing diseases in horseradish. In the present study, 89 Alternaria spp. isolates from Armoracia rusticana, sampled from nine districts in Serbia, were characterized based on their morphology, physiology, and molecular markers. Morphological characterization and molecular analyses based on ITS, GAPDH, Alt a 1, and PM-ATP sequences identified three distinct species associated with the disease: Alternaria brassicae, A. brassicicola, and A. alternata. For all species, growth and sporulation rates at 0, 5, 10, 15, 20, 25, 30, 35, and 40°C showed a quadratic response, with A. alternata having the widest temperature optimum (20 to 30°C) while A. brassicicola had higher optimum temperatures (20 to 25°C) than A. brassicae (15 to 20°C). To gain a better understanding of the pathogenicity of these species, the influence of leaf age, host susceptibility, and ability to infect artificially wounded and nonwounded leaves were tested. The pathogenicity test identified A. brassicicola and A. brassicae as the main causal agents of horseradish leaf spot disease. Results indicated that young and intact leaves of horseradish and cabbage were less susceptible to infection and also suggested the potential for cross-infection between these two hosts. Haplotype networks showed haplotype uniformity for A. brassicae, two haplotype groups of A. brassicicola, and eight haplotype groups of A. alternata in Serbia and suggest the possible association of some haplotypes with the geographic area. This study is the first to investigate Alternaria leaf spot disease on A. rusticana in Serbia and is the first record of A. brassicicola on horseradish in this country.

Melampsora epitea, known to cause rust on the complex of Salix retusa prostrate willows distributed in the subalpine and alpine belt of the mountains of Central Europe, is here documented for the first time in Montenegro and North Macedonia growing at six localities. It is not new for Serbia, but the records come from a newly reported host, namely Salix serpyllifolia. The pathogen?s distribution presumably is wider than initially believed, and further surveys need to be conducted.

Significant percent of world cultural heritage artifacts is threatened by fungal infestation. Fungi can deteriorate different substrates via various physical and chemical mechanisms. Hyphal growth and penetration into the substrate can cause symptoms like discoloration, biopitting, cracking, exfoliation and patina formation. On the other hand, chemical mechanisms include acid secretion, release of extracellular enzymes, pigment production, oxidation/reduction reactions and secondary mycogenic minerals formation. These processes can lead to serious, both esthetic and structural, alterations which may be irreversible and could permanently impair artworks. Proper isolation and identification of autochthonous isolates, as well as employment of different microscopic techniques and in vitro biodegradation tests are pivotal in understanding complex biodeterioration mechanisms caused by microorganisms, including fungal deteriogens. Biodeterioration and biodegradation studies require multidisciplinary approach and close collaboration of microbiologists, chemists, geologists and different personnel responsible for the safeguarding of cultural heritage monuments and artifacts, especially restorers and conservators.

The global demand for rapeseed makes it one of the fastest growing markets in crop production, with a need for increasing growing area and productivity, both of which depend on effective pathogen control strategies. Alternaria pathogens cause serious losses of brassica crops and occur in most rapeseed‐growing regions around the world. In this study, morphological, molecular, and pathogenic analyses of 113 isolates collected from nine important rapeseed‐growing areas in Serbia identified four pathogens: Alternaria brassicae, A. brassicicola, A. japonica, and A. alternata, causing leaf spot disease. Molecular analyses of ITS, GAPDH, Alt a1, and ATP sequences revealed one multilocus haplotype for A. brassicae and A. japonica isolates, whereas for A. brassicicola and A. alternata three and five haplotypes were distinguished, respectively. Pathogenicity tests showed that A. brassicicola was the most virulent while A. brassicae and A. japonica exhibited the same level of pathogenicity. The A. alternata population was generally weakly pathogenic with one nonpathogenic, genetically separated but closely related group of isolates, suggesting that pathogenicity is more unstable in this phylogenetic lineage. The data recorded on rate of growth and sporulation of isolates at 0, 5, 10, 15, 20, 25, 30, 35, and 40 °C revealed significant differences in evolutionary strategies among species, as A. alternata had the widest optimum range and the fastest growth rate, A. brassicicola showed the highest sporulation intensity, and A. brassicae expressed lower optimum temperatures for sporulation compared to other groups. All species indicated the potential for cross‐infection of cabbage, and some haplotypes of A. brassicicola were previously isolated from horseradish in Serbia, suggesting the presence of one persistent Alternaria population on multiple brassica hosts in the region. This report describes the first detailed study of Alternaria spp. in rapeseed in Serbia.

Talaromyces minioluteus is one of the important species of genus Talaromyces, which has cosmopolitan distribution and is encountered on a wide range of different habitats. This species has not been considered as an important plant pathogen, even though it has been isolated from various plant hosts. Fruits and vegetables with Penicillium-like mold symptoms were collected from 2015 to 2017 from markets in Serbia. Isolates originating from quince, tomato, and orange fruits, onion bulbs, and potato tubers were identified and characterized on a morphological, physiological, and molecular level. Morphological and physiological examination included observing micromorphology, testing growth on six different media and at five different temperatures, and production of three enzymes. Molecular identification and characterization were performed using four molecular markers: internal transcribed spacer, β-tubulin, calmodulin, and DNA-dependent RNA polymerase II second largest subunit. The results of morphological and molecular analyses were in agreement, and they proved that the obtained isolates are T. minioluteus. In the pathogenicity assay, T. minioluteus was confirmed as a pathogen of all species tested with the exception of potato tubers. This is the first report of T. minioluteus as a postharvest plant pathogen on quince, tomato, and orange fruit and onion bulbs. Also, this is the first record of T. minioluteus in Serbia.

Paper description: Lamium purpureum L. (Lamiaceae) glandular trichomes and the composition of essential oil were analyzed. Two types of trichomes, non-glandular and glandular (peltate and capitate) were identified. The main compounds of essential oil were trans -anethole (59.61%), fenchone (6.54%), α -pinene (6.14%) and β -pinene (4.98%). The antioxidant activities of the ethanolic plant extracts were determined using DPPH, ABTS, FRAP and TRP assays. Antifungal activity of the ethanolic extract was observed, While the extracts did not exhibit fungicidal activity, morphophysiological changes (sporulation loss, depigmentation of conidial heads) were observed in all tested isolates. L. purpureum could be a new potential source of natural antioxidants. Abstract : The aim of this study was to investigate the structure of glandular trichomes and essential oil composition of Lamium purpureum L. (Lamiaceae) and to determine potential biological activities of its different extracts. The micromorphology and distribution of trichomes on vegetative plant parts were investigated using scanning electron microscopy (SEM). Two types of trichomes were identified: non-glandular (simple, uniseriate, multicellular) and glandular (both peltate and capitate). The essential oil of L. purpureum was isolated by hydrodistillation and analyzed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The main compounds were trans -anethole (59.61%), fenchone (6.54%), α -pinene (6.14%) and β -pinene (4.98%). The total phenol and flavonoid contents of 10%, 30% and 50% ethanol extracts were determined spectrophotometrically. The free radical scavenging and reducing power activity of extracts were analyzed using the DPPH and ABTS radical scavenging methods, and the ferric reducing antioxidant power (FRAP) and total reducing power (TRP) assays. The 50% ethanolic extract exhibited the strongest antioxidative activity while the lowest was determined for the 10% ethanolic extract. Antifungal activity of L. purpureum hydroalcoholic extracts was evaluated against eight Aspergillus species by the microdilution method. Light microscopy (LM) demonstrated structural abnormalities of the tested Aspergilli . While the plant extracts did not exhibit fungicidal activity, morphophysiological changes (sporulation loss and depigmentation of conidial heads) were observed in all tested isolates. https://doi.org/10.2298/ABS200117019G Received: January 17, 2020; Revised: February 25; Accepted: April 7, 2020; Published online: April 23, 2020 How to cite this article: Grujic SM, Savkovic ŽD, Ristic MS, Džamic AM, Ljaljevic Grbic MV, Vukojevic JB, MarinPD. Glandular trichomes, essential oil composition, anti- Aspergillus and antioxidative activities of Lamium purpureum L. ethanolic extracts. Arch Biol Sci. 2020;72(2):253-63.

Neurological disorders, especially Alzheimer’s and Parkinson’s diseases, are among the leading causes of death and disability nowdays. Ganoderma lucidum and Salvia officinalis have been used for prevention and treatment of numerous health problems since ancient times. However, despite the fact that their single extracts possess very strong antioxidant and antineurodegenerative properties, the effect of their combination has not yet been examined. Accordingly, the aim of this study was to investigate possible synergistic effects of combined G. lucidum and S. officinalis ethanol and aqueous extracts. The combinations were obtained by extraction of mixed G. lucidum and S. officinalis dried materials, as well as by mixing of their extracts in different proportions (70:30, 50:50, 30:70). Generally, S. officinalis extracts were stronger antioxidants than those of G. lucidum, and were especially more efficient reducers of Fe3+ and neutralizers of DPPH and ABTS radicals. In DPPH· and ABTS· neutralization, strong synergism was noted for the ethanol extract of a combination of dried G. lucidum and S. officinalis materials (30:70). With respect to acetylcholinesterase inhibition, strong synergism was noted for the mixture of dried G. lucidum and S. officinalis materials (70:30) and mixture of their extracts (30:70), while the same effect on tyrosinase activity was observed only in the case of a mixture of equal dried materials. It follows that efficient G. lucidum/S. officinalis mixtures could be a basis for producing novel food ingredients with antioxidant and antineurodegenerative properties.

The goals of the present study were to characterize the profile of ligninolytic enzymes in five Pleurotus species and determine their ability to delignify eight common agro-forestry residues. Generally, corn stalks were the optimal inducer of Mn-dependent peroxidase activity, but the activity peak was noted after wheat straw fermentation by P. eryngii (3066.92 U/L). P. florida was the best producer of versatile peroxidase, especially on wheat straw (3028.41 U/L), while apple sawdust induced the highest level of laccase activity in P. ostreatus (49601.82 U/L). Efficiency of the studied enzymes was expressed in terms of substrate dry matter loss, which was more substrate-than species-dependent. Reduction of substrate dry mass ranged between 24.83% in wheat straw and 8.83% in plum sawdust as a result of fermentation with P. florida and P. pulmonarius, respectively. The extent of delignification of the studied substrates was different, ranging from 51.97% after wheat straw fermentation by P. pulmonarius to 4.18% in grapevine sawdust fermented by P. ostreatus. P. pulmonarius was also characterized by the highest cellulose enrichment (6.54) and P. ostreatus by very low one (1.55). The tested biomass is a highly abundant but underutilized source of numerous value-added products, and a cocktail of ligninolytic enzymes of Pleurotus spp. could be useful for its environmentally and economically friendly transformation.

The aim of the study was comparative analysis of degradation of wheat straw lignin by white-rot fungi and its implications on the efficiency of enzymatic hydrolysis of holocellulose. Cyclocybe cylindracea, Ganoderma resinaceum, Irpex lacteus, Pleurotus ostreatus and Trametes versicolor were the species studied. Peroxidases were predominantly responsible for lignin degradation even though high laccase activities were detected, except in the case of Irpex lacteus where laccase activity was not detected. Studied fungal species showed various ability to degrade lignin in wheat straw which further affected release of reducing sugars during enzymatic saccharification. The highest rate of lignin degradation was noticed in sample pretreated with Irpex lacteus (50.9 ± 4.1%). Among all tested species only Ganoderma resinaceum was suitable lignin degrader with the 2-fold higher hydrolysis yield (51.1 ± 4.7%) than in the control, and could have significant biotechnological application due to lower cellulose loss. A key mechanism of carbohydrate component convertibility enhancement was lignin removal in the biomass. Long time consumption, the low sugar yields and unpredictable fungal response still remain the challenge of the fungal pretreatment process.

Previous studies showed that some mushrooms are highly efficient in inhibiting acetylcholinesterase and tyrosinase, the increased activity of which can trigger the development of Alzheimer and Parkinson diseases. Starting from the fact that free radicals at high concentrations could cause neurodegenerative disorders as well as great interest in new, natural antineurodegenerative drugs, the goal of this study was to determine the in vitro antioxidative and neuroprotective potentials of various Pleurotus ostreatus and Laetiporus sulphureus extracts. L. sulphureus was a better antioxidative agent; it showed higher reducing power, was a more efficient scavenger of DPPH and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radicals, and was an Fe3+ reducer. The most efficient acetylcholinesterase inhibitor was hot water extract of P. ostreatus fruiting body, which was slightly weaker than the commercial preparation, galantamine. However, in comparison with α-kojic acid, tested extracts were weaker tyrosinase inhibitors. Considering that tested extracts were rich in phenols and that their amounts were in positive correlation with the extent of radical neutralization and acetylcholinesterase and tyrosinase inhibition, it is assumed that these compounds are the potential carriers of the neuroprotective activities. Owing to the significant antioxidative and antineurodegenerative capacity of these species, they can be suggested as novel nutraceuticals and pharmaceuticals.

The main purpose of this study was to isolate airborne fungi and assess seasonal variations in air contamination with their particulates by determining the levels of their propagules in the nave and exonarthex of a church. We also monitored indoor microclimate as a determining factor for fungal proliferation on wall paintings, spore release, and transmission through the air. The temperature and relative humidity of the nave favoured fungal growth. A total of 33 fungi were isolated, mainly of the phylum Ascomycota, and to the lesser extent of the phyla Zygomycota and Basidiomycota. The most common were the fungi of the genera Penicillium and Aspergillus (23.55 % and 20.58 %, respectively). Sørensen’s quotient of similarity (0.37) suggests moderate species overlap and constant exchange of fungal propagules between the nave and exonarthex. The autumn had the highest diversity, with 17 documented taxa, followed by the summer and the winter. The spring had only eight taxa. Quantitative analysis of the airborne mycobiota in the nave (430±84.85 to 1880±106.07 CFU m-3) and exonarthex (715±59.62 to 2295±91.92 CFU m-3) showed very high contamination throughout the year, with values exceeding the maximum permissible concentrations by most standards. Many of the fungi determined in this study are known for their biodeteriogenic, toxigenic, and allergenic properties, and are a threat not only to occasional visitors and staff, but also to valuable works of art decorating nave walls., Glavni cilj ovoga istraživanja bio je izolirati gljive iz zraka i procijeniti sezonske promjene u onečišćenju zraka gljivičnim propagulama u naosu i egzonarteksu istraživane crkve. Također su praćeni mikroklimatski parametri kao ograničavajući čimbenici za razvoj i rast gljiva na zidnim slikama, za oslobađanje spora i njihovu transmisiju kroz zrak. Zabilježena temperatura i relativna vlažnost zraka u naosu pogodovale su razvoju i rastu gljiva. Dokumentirana su 33 taksona gljiva, uglavnom pripadnika koljena Ascomycota, a manje su zastupljeni oni Zygomycota i Basidiomycota. Najčešće su bile prisutne plijesni rodova Penicillium (23,55 %) i Aspergillus (20,58 %). Sørensenov indeks sličnosti (0,37) upućuje na stalnu i umjerenu razmjenu gljivičnih propagula između naosa i egzonarteksa. Uzorci uzeti u jesen pokazali su najveću raznolikost sa 17 zabilježenih taksona, a oni uzeti u proljeće samo osam taksona. Kvantitativna mikološka analiza u naosu (430±84,85 do 1880±106,07 CFU m-3) i egzonarteksu (715±59,62 do 2295±91,92 CFU m-3) pokazala je visoku godišnju razinu onečišćenja zraka, s vrijednostima koje prema većini standarda prelaze dopuštene koncentracije. Mnoge identificirane gljive mogu dovesti do biodeterioracije, proizvesti mikotoksine i izazvati alergijske reakcije. Stoga su nepoželjne u sakralnim objektima ne samo zbog očuvanja murala nego i zbog zaštite zdravlja zaposlenih, posjetitelja i konzervatora.

This study was aimed to analyze the chemical composition and biological activities of extracts of Salvia fruticosa and S. lanigera originated from Libya. Dichloromethane, ethyl acetate, methanol, ethanol and water extracts obtained from wild growing plants were analyzed for the composition using HPLC-DAD, which revealed presence of phenolic acids and flavonoids, mainly in alcoholic and water extracts. Total phenolic and flavonoid contents, as well as anti-oxidant activity using 2.2-diphenyl-1-picrylhydrazyl (DPPH), 2.2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid diammonium salt (ABTS), ferric reducing anti-oxidant potential (FRAP) and β-carotene bleaching (β-CB) assays were determined spectrophotometrically. Extracts of S. fruticosa exhibited stronger activity in all applied assays, especially ethanol extract in DPPH assay (IC50, 29.55 μg/mL) and in β-CB assay (85.11%). The ethanol and water extracts were selected for further investigation, because of their wide usage in phytotherapy. The extracts were screened for the antimicrobial activity against 11 bacteria (six Gram-negative and five Gram-positive) and seven fungi using microdillution method. Ethanol extracts showed stronger activity than water extracts, particularly against Gram-positive bacteria. Trichophyton mentagrophytes was the most sensitive to the water extract of S. lanigera (MIC, minimal inhibitory concentration and MFC, minimal fungicidal concentration, 8 mg/mL). Cytotoxic activity on human carcinoma cell line HCT-116 was determined by 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyl tetrazolium bromide (MTT) cell viability assay, where only ethanol extract of S. fruticosa demonstrated certain activity (IC50, 375.96 μg/mL). The testing of anti-neurodegenerative activities of extracts showed better tyrosinase inhibiting effect (55.26–74.66%) than standard kojic acid (33.93–51.81%), while the extracts were less effective against acetylcholinesterase compared with standard galanthamine. According to the obtained results, S. fruticosa and S. lanigera originated from Libya have proved to be the promising source of natural compounds possessing a range of biological activities.

Background Microfungi are causal agents of numerous diseases and disorders of agricultural plants, farm mushrooms and animals as well as human, which results are serious global reduction of the food amount, decrease of life quality, the severe life-threatening diseases and enormous economic losses. Methods In spite of organism innate ability to combat against pathogens, in invasions of some pathogens, support of additional antimycotic agents to defence system is required. Nowadays, common "fighters" against the microfungi are numerous synthetic fungicides that, besides benefits, have also side effects on host and environment and can cause the development of fungicide resistance in the pathogens. Therefore, the creation of new natural fungicides with different modes of action, strengthening the defense system and increase of organism resistance to pathogens are the main requirements of modern society. Results Numerous mushrooms produce chemically various intra- and extracellular metabolites with antifungal potential, among which the most potent ones are polysaccharides, proteins, and phenolic compounds. They act as immunostimulators, inhibitors of pathogen development and virulence and/or activators of pathogens` autolytic system. Conclusion Therefore, mushroom-based antimycotic agents could be successfully applied in the diseases treatments as accessories or alternatives to commercial therapies and in such a way contribute to environmentally friendly combat against pathogens, i.e. decrease or complete substitution of commercial synthetic fungicides with natural ones.

Demand for Ganoderma lucidum basidiocarps on the global market is increased due to its numerous health benefits. As they are rare in nature and traditional cultivation on logs is not ecologically and economically justified method, current trend is finding of good alternative substrate for production. Whether wheat straw, the most abundant crop residue in Europe, could be a novel substrate for G. lucidum cultivation was the question which led to the definition of the goals. Two wild and one commercial strains were objects of the study. Despite some morphological differences among basidiocarps, all strains belong to G. lucidum sensu stricto, which was confirmed by analyses of ITS, tef1-a and rpb2 gene sequences. Wheat straw showed as a good substrate, namely the periods required for the complete colonisation of wheat straw by mycelium as well as the formation of primordia and basidiocarps were relatively short. The totally fresh and dry yield and biological efficiency were also significant. If it is taken into consideration that untreated wheat straw was used, validities of its introduction in industrial-scale G. lucidum cultivation can be demonstrated.

The aim of this study was to optimize cultivation conditions for Mn-oxidizing peroxidases and laccase production and selective degradation of beech wood and wheat straw lignin by Trametes gibbosa. To promote lignin degradation, the effect of different carbon and nitrogen sources, type of cultivation, enzyme dynamics and inducers were studied. Solid-state cultivation was optimal for lignin degradation. Wheat straw was the optimal carbon source for peroxidase activities stimulating the synthesis of numerous isoforms. The dynamics of enzymatic activities showed that 19-day-old fermentation is optimal and that activities were directly associated with enzyme production. Alteration of nitrogen sources and inducers was applied to increase the rate of lignin degradation and decrease cellulose degradation. The presence of nitrogen in the form of (NH4)(2)SO4 and concentration of 10 mM significantly increased the extent and selectivity of delignification of wheat straw compared with cellulose (44.1 vs. 36.1%). The most effective and selective wheat straw degradation (52.0% of lignin vs. 31.3% of cellulose) was achieved by enriching the optimum medium, as defined above, with 1.0 mM p-anisidine. The optimum conditions for wheat straw processing to achieve delignification and production of highly active ligninolytic enzymes by T. gibbosa were (NH4)(2)SO4/wheat bran medium supplemented with p-anisidine. This study indicated the significant potential of optimizing external factors as a promising tool for induction and selectivity in the degradation of lignocelluloses by T. gibbosa as a pretreatment for several biotechnological processes.

Leaf spot diseases caused by

Bat guano is an important source of microbial diversity in caves and can be a source of potential pathogens. Laemostenus (Pristonychus) punctatus is a guanophilic ground beetle species, which pygidial gland secretion exhibits action against pathogenic and other microbes. The distribution and diversity of microbes in bat guano from a karstic cave were determined in this study. Additionally, antimicrobial activity of the pygidial gland secretion of L. (P.) punctatus against guano-dwelling microbes was tested; minimal inhibitory concentration (MIC) and chemical composition of the secretion were analyzed. In total, 63 different bacterial species and 16 fungal morphotypes were isolated from guano samples by the cultivation method and confirmed using and phenotypic characterization and molecular identification. There was a difference in the composition of certain microorganisms between the sampling points (cave locations) and between the guano layers. The largest number of bacterial isolates belongs to the genera Lysinibacillus and Paenibacillus, while Pseudomonas species were highly abundant at the innermost sampling point. For the guanophilic fungi, the majority are ascomycetes, with Penicillium and Aspergillus as the most dominant genera. Meyerozyma guilliermondii was the only yeast species found in the guano samples. The most sensitive isolates were Enterococcus eurekensis (MIC 0.007 mg/mL) and Escherichia fergusonii (MIC 0.028 mg/mL). The most sensitive fungal isolates were M. guilliermondii, Penicillium expansum, and Trichoderma harzianum (MIC 0.15 mg/mL). This study opens a new possibility for better understanding of ecological relations between microorganisms and troglophilic ground beetles and for detailed investigations of morpho-anatomical aspects of pygidial glands.

Ganoderma lucidum has a long tradition of use in folk medicine of the Far East, which is documented in the oldest Chinese pharmacopoeia, written in the first century B.C, declaring it a superior medicine. The healing properties of G. lucidum reflected on folk names such as: Reishi, Mannentake, Ling Zhi etc., which mean ?herb of spiritual power?, ?mushroom of immortality? or ?10,000-year mushroom?, respectively. It has been known, for thousands of years, that this species extends life span, increases youthful vigour and vitality and it was used in the treatments of hepatitis, kidneys? disease, hypertension, arthritis, asthma, bronchitis, arteriosclerosis, ulcers and various types of cancer. However, Western civilisation did not discover its healing properties until the 20th century. Modern scientific researches and numerous clinical trails, conducted in recent decades, have confirmed the ancient knowledge of Eastern nations and given them a scientific basis. These studies have demonstrated many biological activities of G. lucidum extracts and compounds, including: immunomodulating, antioxidative, cytotoxic, hypoglycaemic, anti-inflammatory, antiallergic, antimicrobial, etc. It has been reported that its extracts play important role in detoxification of the body and protecton of the liver, as well as in reducing cardiovascular problems, stress and anxiety. However, its most important effect is undoubtedly immunostimulating one as it is the basis of many other positive effects. The Japanese government introduced G. lucidum on the official list of auxiliary agents for the treatments of various cancers, Alzheimer?s disease, diabetes and chronic bronchitis. Many chemical components have been isolated from G. lucidum, but polysaccharides and terpenoids are the main carriers of its bioactivities.

Although some members of the genus Daedaleopsis have been prized for their medicinal and spiritual powers since Neolithic times, modern science has not yet sufficiently dealt with their bioactivities. This study aims at defining the antioxidative ac­tivities of extracts of Daedaleopsis tricolor wild and cultivated basidiocarps and mycelium and assessing their dependance on substrate type. Ethanol extracts (at a concentrations from 0.25 mg/mL to 16.00 mg/mL) of mycelium and wild fruiting bodies showed a considerable antioxidative potential (88.65% and 81.57%, respectively), which was almost the same as the commercial antioxidant BHA (88.91%). These radical scavenging abilities were reflected in EC50 values, which were 12.45 mg/mL for the extract of cultivated basidiocarps, 8.29 mg/mL for the extract of wild basidiocarps, 7.93 mg/mL for mycelium one, and 0.10 mg/mL for commercial antioxidant. Despite the fact that phenol proportion in the extracts was no neg­ligible (between 20.41 μg GAE/mg of the extract of dry wild basidiocarps and 146.37 μg GAE/mg of the extract of dry cultivated basidiocarps), its correlation with antioxidative activity was moderate. Flavonoids, in significant concentration, were detected only in the extract of cultivated fruiting bodies (28.64 μg QE/mg of dry extract), but no correlation with radical scavenging capacity was noted. A remarkable antioxidant potential, especially of the submerged cultivated mycelium, put D. tricolor high on the list of promising new natural antioxidants. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 173032]

Biodeterioration caused by fungal colonizers on an ancient stone stela, excavated from the former Roman settlement (Eastern Serbia) was investigated. According to selected biodeterioration elements, average deterioration index was assessed (0.8), prompting the need for conservation. Fungal somatic and reproductive structures, along with lichen thalli and moss fragments, were detected on the surface using different microscopy and cultivation methods. In situ microscopy on the site was implemented, for the first time, in the study of stone monuments. Biodeteriorative potential of 5 selected isolates was tested using qualitative biochemical tests, SEM-EDS and XRPD analyses. Fusarium proliferatum and Penicillium crustosum altered the pH value in broth minimal medium. Pigment production was demonstrated for F . proliferatum , while P. crustosum showed potential for calcite dissolution. All isolates induced biomineralization on solid medium with calcium acetate, where weddellite, calcite and subordinate whewellite crystals were confirmed via SEM-EDS and XRPD. Weddellite and calcite production was documented for P. crustosum in solid medium with calcium carbonate. Conservation treatment was carried out with benzalkonium chloride-based biocide, in addition to mechanical treatment. After conservation, in situ microscopy showed deteriorated stone surface covered with residual lichen thalli fragments, while mycelium and reproductive structures of micromycetes were not detected.

Ganoderma applanatum is a widely-distributed wood decaying species whose ligninolytic enzyme system has not been sufficiently studied. The aims of the study were to profile its Mn-oxidizing peroxidases and laccases and defining of wheat straw and oak sawdust delignification extents depending on cultivation type. Activities of these enzymes were higher in submerged than in solid-state cultivation. Oak sawdust induced the highest activities of Mn-dependent (5545.5 U L −1 ) and Mn-independent peroxidases (5810.0 U L −1 ), and wheat straw stimulated laccase activity (11007.0 U L −1 ). The isoelectric focusing profiles of enzymes and extent of lignocellulose degradation were affected by plant residue as well as type and period of cultivation. Submerged cultivation induced the synthesis of a higher number of enzyme isoforms and the maximum levels of lignin, hemicellulose and cellulose degradation (40.9%, 32.7% and 27.4%) were reached during this oak sawdust fermentation. However, selectivity in fiber mineralisations was the highest during solid-state fermentation of wheat straw, which is important for possible application in various biotechnological processes that require accessible cellulose, such as production of more digestible feed, paper pulp and bioethanol.

An extensive aeromycological survey was performed in the premises of a cultural heritage conservation facility to estimate fungal propagule concentrations in both indoor and outdoor air and seasonal dynamics with regard to temperature and air humidity. The highest average propagule concentration in indoor air was documented in spring, and for outdoor air, in summer. Increased propagule loads were documented in most rooms during winter, fall and spring, and in all examined rooms in summer. Very high propagule concentrations (>25,000 CFU m−3) were reported in ground-floor rooms. The total mycobiota was comprised of 74 fungal species, with a prevalence of Aspergilli and Penicillia. Cladosporium and Penicillium isolates were found to be the most abundant in all seasons in both indoor and outdoor air samples. Among the identified fungi, potential human pathogens, allergens and mycotoxin producers were present. Numerous documented species play an important role in the deterioration of cultural heritage artifacts and are known producers of extracellular enzymes, acids and pigments. Application of a selective medium (M40Y) allowed isolation of various xerophiles and xerotolerant species. Aeromycological analyses are mandatory for determining the appropriate conditions for the protection of the health of conservators, as well as the objects of cultural heritage.

The study addresses in vitro degradation potential of airborne Aspergillus and Penicillium/Talarmyces species originating from cultural heritage conservation premises. A series of rapid, cost effective biodegradation assays were performed to assess production of extracellular pigments, acids, and enzymes. Most of the isolates have demonstrated positive growth in at least one of the preformed tests. Strongest overall degradation potential was demonstrated for Penicillium brevicompactum, P. glabrum, and Talaromyces sayulitensis while Aspergillus domesticus, A. penicillioides, A. pseudoglaucus, and A. ruber did not exhibit positive reaction in any of the employed assays. Majority of isolates exhibited proteolytic and cellulolytic activity while carbonate dissolution was observed for only five tested fungi. Highest alteration of pH value in liquid media was documented for T. sayulitensis while A. niger and P. expansum exhibited strongest acid production on CREA. Certain isolates, mostly Penicillium species, displayed production of extracellular pigments. The results imply that many of the tested fungi have significant biodegradation capacity, indicating their potential to inflict structural and esthetic alterations on cultural heritage objects.

Cancer is the second cause of morbidity and mortality worldwide, i.e., half of the men and more than a third of women of the world population get sick with some type of cancer during a lifetime, and one-quarter of all adults die of this disease. Common treatments, chemo- and radiotherapy, are not highly effective, give satisfactory results only in the treatment of early cancer development stages or have no any effect on some cancer types, and commonly cause numerous side effects. Therefore, alternative medicine based on various natural sources attracts great attention nowadays. Although mushrooms, their extracts, and isolated metabolites cannot be considered drugs, they are a type of important dietary supplement, i.e., functional food or nutraceuticals, and could be used as auxiliary natural cytostatics. They are highly selective, i.e., not toxic or almost nontoxic to normal cells, do not cause any side effects, and even reduce harmful effects caused by conventional treatments, and finally, resistance to them cannot be developed. Mushroom extracts or biologically active compounds isolated from them affect cytotoxic activity on a few mechanisms: stimulation of immune system; antioxidative, antimutagenic, and anti-inflammatory activity; regulation of expression of regulators of some cell processes; cell cycle arrest and apoptosis; disturbance of DNA synthesis and structure; changes in morphology and mobility of malignant cells; and antiangiogenic activity.

Members of the millipede order Julida rely on dominantly quinonic defensive secretions with several minor, non-quinonic components. The free radical-scavenging activities of ethanol, methanol, hexane, and dichloromethane extracts of defensive secretions emitted by Pachyiulus hungaricus (Karsch, 1881) and Megaphyllum unilineatum (C. L. Koch, 1838) were investigated using the ABTS, DPPH, and total reducing power (TRP) tests. The obtained extracts were also tested for inhibition of acetylcholinesterase and tyrosinase activity. Finally, the antifungal potential of both julid extracts was evaluated against seven Fusarium species. Secretions of both species showed activity against free radicals, acetylcholinesterase, tyrosinase, and all of the selected fungal species. The secretions of P. hungaricus exhibited a more potent antioxidative effect than did those of M. unilineatum, while there were no significant differences of antiacetylcholinesterase activity between the tested extracts. Only the hexane extract of M. unilineatum showed an effect on tyrosinase activity stronger than that of P. hungaricus. Fusarium sporotrichioides, F. graminearum, and F. verticillioides were the fungi most resistant to secretions of both julids. The Fusarium species most susceptible to the secretion of P. hungaricus was F. avenaceum, while the concentrations of M. unilienatum extracts needed to inhibit and completely suppress fungal growth were lowest in the case of their action on F. lateritium. Our data support previous findings that julid defensive secretions possess an antimicrobial potential and reveal their antioxidative and antineurodegenrative properties. Bearing in mind the chemical complexity of the tested defensive secretions, we presume that they can also exhibit other biological activities.

Millipedes use an array of chemical compounds to defend themselves from predator attack. These chemical substances can have additional roles, i.e. defence against various pathogens. We evaluated the efficacy of the defensive secretion of Apfelbeckia insculpta (L. Koch, 1867) against bacteria, yeasts, and filamentous fungi. The tested secretion consisted of two compounds, p-cresol and phenol, and showed antibacterial, antibiofilm, and antifungal potential against all selected microorganisms. The most sensitive bacterium in our study was Pseudomonas aeruginosa, while the tested defensive secretion manifested the lowest activity against Escherichia coli. The defensive secretion of A. insculpta also showed an ability, albeit mild, to suppress biofilm formation by P. aeruginosa. Among the tested yeasts, Candida albicans and C. krusei were the most susceptible and most resistant species, respectively. Finally, the concentration of extracts obtained from the tested defensive secretion needed to achieve an antifungal effect was lowest in the case of Cladosporium cladosporioides. Fusarium verticillioides and Penicillium rubens were the micromycetes most resistant to the tested secretion. Our results indicate that antibacterial activity of the defensive secretion of A. insculpta is similar to or slightly weaker than that of streptomycin, while comparison with antimycotics showed that the tested millipede secretion has stronger activity than fluconazole, but weaker activity than nystatin and ketoconazole. The present study corroborates previous findings indicating that the defensive secretions of millipedes can have different roles apart from antipredator protection and are effective against pathogenic microorganisms.

The antifungal activities of Origanum vulgare essential oil (EO) and of a biocide based on silver and hydrogen peroxide (Sanosil S003) against seven Aspergillus species isolated from different substrata (stone, brick, silk and paper) of cultural heritage objects in Serbia were evaluated. Microdilution, agar dilution and microatmosphere methods were used to determine minimal fungistatic and minimal fungicidal concentrations (MIC and MFC), and light microscopy to determine structural abnormalities. MIC and MFC values for O. vulgare EO ranged from 0.2 to 5 mg mL−1and for Sanosil S003 from 5 to 250 mg mL−1. Aspergillus sp. sect. fumigati was the most susceptible isolate, where MIC and MFC values were achieved at 0.5 mg mL−1for O. vulgare EO, while MIC and MFC values for Sanosil S003 were achieved at 5 and 10 mg mL−1, respectively. Morpho-physiological changes were documented in all isolates, including lack of sporulation, depigmentation of conidiogenous apparatus and conidia, and presence of aberrant fungal structures. O. vulgare EO exhibited stronger anti-Aspergillus activity than Sanosil S003, as demonstrated by the higher MIC and MFC values and fewer morpho-physiological changes observed in the tested Sanosil S003 concentrations. O. vulgare EO could be an excellent alternative to commercial biocides, with high potential in the field of cultural heritage conservation.

The study aimed to evaluate the antiradical and antimicrobial (antibacterial and antifungal) potentials of ethanol mycelial extracts of selected Ganoderma species and strains and to define interand intraspecies diversity among Ganoderma species and strains. Ganoderma lucidum strains were good DPPH• scavengers (neutralizing up to 57.12% radicals), contrary to G. applanatum (20.35%) and G. carnosum (17.04%). High correlations between the activities and contents of total phenols in the extracts showed that these compounds were carriers of the activity. Results obtained by both discdiffusion and microdilution methods indicated that the extract of G. lucidum BEOFB 433 was the most potent antibacterial agent that inhibited growth of almost all bacterial species at a concentration of 1.0 mg/mL. Salmonella typhimurium was the most sensitive species to the mycelium extracts. Extracts of G. lucidum BEOFB 431 and BEOFB 434 showed the best antifungal activity since in concentration of 0.5 mg/mL inhibited the growth of Aspergillus glaucus (BEOFB 431) and the growth of A. glaucus and Trichoderma viride (BEOFB 434). Extracts of G. applanatum and G. lucidum BEOFB 431 had the strongest fungicidal effects, with lethal outcomes for A. glaucus and T. viride, respectively, being noted at a concentration of 1.17 mg/mL. Aspergillus niger was proved as the most resistant species.

Ganoderma lucidum is traditionally used in Eastern medicine to preserve vitality, promote longevity, and treat disease. It possesses immunomodulatory, antitumor, antimicrobial, and antiaging activities, among others, but one of the most important is its antioxidant property, which is the basis for other effects, because free radicals trigger many diseases. The substrate commonly used for commercial cultivation of G. lucidum is not environmentally friendly nor economically justified, so there is a need to find new alternative substrates. The aim of this study was to analyze the effect of substrate composition on the bioactivity of G. lucidum basidiocarps. G. lucidum was cultivated on 2 different substrates: (1) a mixture of wheat straw, grapevine branches, and wheat bran, and (2) wheat straw. Commercial fruiting bodies, cultivated on oak sawdust, were used as the control. 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability, total phenols, and flavonoid content were determined spectrophotometrically to define the antioxidative potential of basidiocarp extracts. The comet test was performed to detect the degree of DNA damage in the cells that were exposed to G. lucidum extracts before and after the effect of oxidants. Higher antioxidative potential was observed for the extract of G. lucidum basidiocarps cultivated on wheat straw compared with that from the mixed substrate and especially with commercial ones. The alternatively cultivated basidiocarps also showed stronger antigenotoxic potential compared with commercial ones. The study showed that fruiting bodies produced on wheat straw, one of the most accessible and cheapest crop residues, are more potent antioxidant and antigenotoxic agents than commercially cultivated ones.

Even though numerous lichen species possess significant medical potentials they are still unexplored, and particularly species and strains originating from Serbia. Therefore, the aim of this study was to evaluate the antioxidative and antimicrobial potential of ethanol extracts of Parmelia saxatilis and Pseudoevernia furfuracea collected in Serbia. The tested extracts were good scavengers of DPPH radicals, with capacities ranging from 14.76% to 79.76% in P. saxatilis and from 21.39% to 90.04% in P. furfuracea. In P. saxatilis level of DPPH• neutralisation was highly correlated with phenol content (r2 = 0.9981) and in P. furfuracea with amount of total flavonoides (r2 = 0.9641). The extract of P. furfuracea inhibited the growth of all tested microorganisms with exception of Aspergillus flavus, while P. saxatilis extract affected only growth of bacterial species. Among tested microorganisms, Staphylococcus aureus and Klebsiella pneumoniae were the most sensitive, while Enterococcus faecalis, Pseudomonas aeruginosa as well as micromycetes were the least sensitive to tested extracts. Because of these potentials and the fact that their long term usage does not have any negative side effects on organism and development of microbial resistance, the extracts could be included in conventional therapy. [Projekat Ministarstva nauke Republike Srbije, br. 173032]

The bioactivity of Ganoderma lucidum basidiocarps has been well documented, but there are no data on the medicinal properties of its submerged cultivation broth nor on the other species of the genus Ganoderma. Thus the aim of this study was to test the potential antimicrobial and antioxidant activity of fermentation broth obtained after submerged cultivation of G. applanatum, G. carnosum, and G. lucidum. DPPH· scavenging ability, total phenols, and flavonoid contents were measured to determine the antioxidative potential of Ganoderma spp. fermentation filtrates, whereas their antimicrobial potential was studied using the microdilution method. DPPH· scavenging activity of G. lucidum fermentation filtrates was significantly higher than that of G. applanatum and G. carnosum, with the maximum (39.67%) obtained from strain BEOFB 432. This filtrate also contained the highest concentrations of phenols (134.89 μg gallic acid equivalents/mL) and flavonoids (42.20 μg quercetin equivalent/mL). High correlations between the activity and phenol content in the extracts showed that these compounds were active components of the antioxidative activity. G. lucidum strain BEOFB 432 was the most effective antibacterial agent, whereas strain BEOFB 434 has proven to be the most effective antifungal agent. The study showed that Ganoderma spp. fermentation filtrates are novel potent antioxidative and antimicrobial agents that could be obtained more quickly and cheaper than basidiocarps.

This comprehensive study was carried out in order to investigate composition and biological activities of essential oil and extracts of Salvia ringens Sibth. & Sm. (Lamiaceae) originating from Macedonia. Major components of the oil, analyzed using GC-FID and GC–MS, were monoterpenes 1.8-cineole (31.99%), camphene (17.06%), borneol (11.94%) and α-pinene (11.52%). HPLC analysis showed presence of 17 phenolic components, mainly in methanol and ethyl acetate, followed by ethanol, water and dichloromethane extracts. Total phenolics and flavonoids as well as DPPH, ABTS, and FRAP activities were measured spectrophotometrically. Essential oil, ethanol, and water extracts showed antimicrobial activity using microdilution method. Ethanol and water extracts performed cytotoxic activity against colon carcinoma HCT-116 cell line using MTT assay. According to the obtained results, S. ringens herb can be considered as the potential source of the essential oil and/or raw material for the extraction and isolation of natural compounds with a range of biological activities.