Caddisflies (Trichoptera) of Protected Calcareous Fen Habitats: Assemblages, Environmental Drivers, Indicator Species, and Conservation Issues.

The caddisflies (Trichoptera) of calcareous fen habitats, in contrast to those of other peatland types, have been poorly researched. We thus conducted a two-year study in south-eastern Poland encompassing four types of such habitats-drained and undrained fens and water bodies (pools and ditches) located within the fens-in order to define trichopteran reference assemblages (PCoA), indicator species (IndVal analysis), and the drivers (both natural and those associated with landscape management, including area protection) responsible for caddisfly species distribution (CCA). The most important environmental driver was habitat persistence. Distance-based RDA analysis revealed a distinct pattern in the distribution of species with or without diapause along the persistence gradient. Environmental drivers associated with plants were also crucial for both fens and water bodies. The key factor influencing the caddisfly assemblages of pools and ditches was the use and management of the surrounding land, whereas in the fens, it was the level of area protection. Physical and chemical water parameters had no statistically significant impact on the assemblages. Some factors can be modified by humans (e.g., water level regulation, vegetation, and landscape management) to maintain healthy ecosystems for aquatic insects.

Microbial Community Response to Various Types of Exogenous Organic Matter Applied to Soil.

Recycling of solid biowaste and manure would reduce the dependence of agriculture on synthetic products. Most of the available studies on the effects of exogenous organic matter (EOM) application to soil were focused on nutrients and crop yield, with much less attention to microbiological processes in soil, especially using modern molecular methods. The aim of this study was to evaluate the effects of various types of manure, sewage sludge and bottom sediment on the biochemical activity and biodiversity of soil and plant yield in a pot experiment. The soil was treated with a range of EOM types: six types of manure (cattle, pig, goat, poultry, rabbit and horse manure; two bottom sediments (from urban and rural systems); and two types of municipal sewage sludge. All EOMs stimulated dehydrogenases activity at a rate of 20 t ha-1. Alkaline phosphatase was mostly stimulated by poultry manure and one of the sludges. In general, the two-fold greater rate of EOMs did not further accelerate the soil enzymes. The functional diversity of the soil microbiome was stimulated the most by cattle and goat manure. EOMs produce a shift in distribution of the most abundant bacterial phyla and additionally introduce exogenous bacterial genera to soil. Poultry and horse manure introduced the greatest number of new genera that were able to survive the strong competition in soil. EOMs differentiated plant growth in our study, which was correlated to the rate of nitrate release to soil. The detailed impacts of particular amendments were EOM-specific, but in general, no harm for microbial parameters was observed for manure and sludge application, regardless of their type. There was also no proof that the PAH and pesticide contents measured in manure or sludge had any effect on microbial activity and diversity.

Differentiation and Propagation Potential of Arnica montana L. Achenes as a Consequence of the Morphological Diversity of Flowers and the Position of Flower Heads on the Plant.

Arnica montana L. is a very important medicinal plant and simultaneously a European endemic endangered plant species. The morphological features and details of seed development and achene variability are poorly recognized. The aim of this study was to determine the impact of the achene position in the infructescence and the location of the inflorescence on the plant on the (i) morphological characteristics and germination ability of achenes, and (ii) recruitment of seedlings and their biometric features. Infructescences containing fully ripe achenes were randomly collected from A. montana individuals for the measurements and the germination experiment. Scanning electron microscopy, fluorescence microscopy, and light microscopy were used for characterization of flowers and achenes. The morphological traits of achenes and reproductive characteristics of A. montana were determined by the position of the achenes in the infructescence and the location of the inflorescence on the plant. The surface of arnica achenes is equipped with non-glandular and glandular trichomes, which is very rarely presented in species of the family Asteraceae. It is possible that the fluid-containing glandular trichomes are a source of essential oils. The peripherally located achenes were longer, thinner, and lighter. They were characterized by lower embryo weight, lower embryo/achene weight ratio, and lower germination capacity in comparison to the centrally located ones. The results presented in this article fill the gap in the knowledge of the morphology of achenes and the biology of the species, and provide information that can help in breeding programs, active protection, and field cultivation.

Variation in Population and Solvents as Factors Determining the Chemical Composition and Antioxidant Potential of Arctostaphylos uva-ursi (L.) Spreng. Leaf Extracts.

The bearberry Arctostaphylos uva-ursi (L.) Spreng. has a long history of ethnopharmacological use. This species has been used in folk medicine for centuries as a rich source of raw material abundant in secondary metabolites and is important for medicinal and pharmacological purposes. The plant is a source of herbal material-Uvae ursi folium, which is highly valued and sought by pharmaceutical and cosmetic industries. The studied bearberry leaves can be classified as a suitable herbal material for use in pharmacy; therefore, the investigated populations can be a potentially valuable source of plant material for cultivation and can be used in in vitro cultures and in biotechnological processes. The objective of this study was to characterize the variability of the phytochemical composition and antioxidant activity of water and ethanol bearberry extracts from raw material collected from different natural populations. In each of the twelve A. uva-ursi sites, three leaf samples were collected and analyzed. The water extracts from bearberry leaves were characterized by similar concentration of arbutin (77.64-105.56 mg g-1) and a significantly higher concentration of hydroquinone (6.96-13.08 mg g-1) and corilagin (0.83-2.12 mg g-1) in comparison with the ethanol extracts -77.21-103.38 mg g-1, 10.55-16.72 mg g-1, 0.20-1.54 mg g-1, respectively. The concentration of other metabolites in the water extracts was significantly lower in comparison with the ethanol extracts. In the case of the water extracts, a significant effect of not only total phenolic compounds, but also hydroquinone on the antioxidant parameters, was observed, which indicates the solvent-related activity of these metabolites. Therefore, it is suggested that special attention should be paid to the concentration of not only arbutin, but also hydroquinone in Uvae ursi folium. The latter metabolite serving a very important function as an active bearberry ingredient should be controlled not only in alcoholic extracts but also in water extracts, since bearberry leaves are applied as infusions and decoctions. The results presented in this paper can contribute to appropriate selection of plant material for pharmaceutical, cosmetic, and food industries, with special emphasis on the antioxidant activity of different types of extracts.

Nitrogen Fertilization and Solvents as Factors Modifying the Antioxidant and Anticancer Potential of Arnica montana L. Flower Head Extracts.

Arnica montana L. is one of Europe's endemic endangered medicinal plants, with diverse biological activities commonly used in medicine, pharmacy, and cosmetics. Its flower heads are a rich source of raw material, with antibacterial, antifungal, antiseptic, anti-inflammatory, antiradical, antioxidant, and antitumor properties. The objective of the present study was (i) to characterize the chemical composition of flower heads of A. montana plants cultivated under nitrogen fertilization, (ii) to identify the impact of the nitrogen fertilization and extraction method (water, ethanol) on the antioxidant activity of extracts, and (iii) to determine the role of different nitrogen doses applied during plant cultivation and different extraction methods in the anticancer activity of the extracts through analysis of apoptosis and autophagy induction in HT29, HeLa, and SW620 cell lines. The present study shows that nitrogen is a crucial determinant of the chemical composition of arnica flower heads and the antioxidant and anticancer activity of the analyzed extracts. Nitrogen fertilization can modify the composition of pharmacologically active substances (sesquiterpene lactones, flavonoids, essential oil) in Arnicae flos. The content of sesquiterpene lactones, flavonoids, and essential oil increased with the increase in the nitrogen doses to 60 kg N ha-1 by 0.66%, 1.45%, and 0.27%, respectively. A further increase in the nitrogen dose resulted in a decrease in the content of the analyzed secondary metabolites. Varied levels of nitrogen application can be regarded as a relevant way to modify the chemical composition of arnica flower heads and to increase the anticancer activity, which was confirmed by the increase in the level of apoptosis with the increase in fertilization to a level of 60 kg N ha-1. The fertilization of arnica plants with low doses of nitrogen (30 and 60 kg N ha-1) significantly increased the LOX inhibition ability of the ethanol extracts. The present study is the first report on the anticancer activity of A. montana water extracts, with emphasis on the role of water as a solvent. In further studies of factors modifying the quality of Arnicae flos, attention should be paid to the simultaneous use of nitrogen and other microelements to achieve synergistic results and to the possibility of a more frequent use of water as a solvent in studies on the biological activity of A. montana extracts.

Chemical Characteristics and Antioxidant Activity of Arctostaphylos uva-ursi L. Spreng. at the Southern Border of the Geographical Range of the Species in Europe.

The bearberry (Arctostaphylos uva-ursi L. Spreng.) is a source of herbal material-bearberry leaf (Uvae ursi folium), which is highly valued and sought by pharmaceutical and cosmetic industries. For many years, leaves of this plant have been used in traditional medicine as a diuretic, antimicrobial, and anti-inflammatory agent for various diseases of the urogenital tract. The bearberry has also been proposed as a natural antioxidant additive due to the high contents of phenolic compounds in its leaves. The study was focused on characterization of the basic phytochemical composition and antioxidant activity of extracts derived from bearberry leaves collected from plants located at the southern border of the geographical range of the species in Europe. The investigated herbal material is characterized by a different chemical profile compared to the chemical profiles of bearberry found in other parts of the continent. Bearberry extracts from plants growing in two different habitat types-heathlands and pine forests showed a wide range of variation, especially in the concentration of hyperoside, corilagin, and methylartutin and the total flavonoid contents. In addition to arbutin, bearberry can be a valuable source of phenolic compounds, which are mainly responsible for the antioxidant properties of extracts. The high content of phenols and high values of antioxidant parameters indicate a high potential of bearberry leaves to be used as a powerful natural source of antioxidants in herbal preparations. Therefore, the A. uva-ursi populations can be a source of plant material for pharmaceutical, cosmetic, and food industries.

Activity and Diversity of Microorganisms in Root Zone of Plant Species Spontaneously Inhabiting Smelter Waste Piles.

The aim was to assess plant driven changes in the activity and diversity of microorganisms in the top layer of the zinc and lead smelter waste piles. The study sites comprised two types (flotation waste-FW and slag waste-SW) of smelter waste deposits in Piekary Slaskie, Poland. Cadmium, zinc, lead, and arsenic contents in these technosols were extremely high. The root zone of 8 spontaneous plant species (FW-Thymus serpyllum, Silene vulgaris, Solidago virgaurea, Echium vulgare, and Rumex acetosa; and SW-Verbascum thapsus; Solidago gigantea, Eupatorium cannabinum) and barren areas of each waste deposit were sampled. We observed a significant difference in microbial characteristics attributed to different plant species. The enzymatic activity was mostly driven by plant-microbial interactions and it was significantly greater in soil affected by plants than in bulk soil. Furthermore, as it was revealed by BIOLOG Ecoplate analysis, microorganisms inhabiting barren areas of the waste piles rely on significantly different sources of carbon than those found in the zone affected by spontaneous plants. Among phyla, Actinobacteriota were the most abundant, contributing to at least 25% of the total abundance. Bacteria belonging to Blastococcus genera were the most abundant with the substantial contribution of Nocardioides and Pseudonocardia, especially in the root zone. The contribution of unclassified bacteria was high-up to 38% of the total abundance. This demonstrates the unique character of bacterial communities in the smelter waste.

Water Soldier Stratiotes aloides L.-Forgotten Famine Plant With Unique Composition and Antioxidant Properties.

Stratiotes aloides L. is common water plant in central Poland. Due to its expansive character, S. aloides L. can strongly affect the functioning of aquatic ecosystems. S. aloides L. was an important famine plant in central Poland. This plant was commonly collected and cooked until the turn of the 20th century. It has also been used to heal wounds, especially when these are made by an iron implement. The objective of the present work was to study the phenolic profile in the leaves and roots of S. aloides as well as their antioxidant potential and ability to inhibit lipoxygenase (LOX) in the light of their potential bioaccessibility. The dominant compound in its leaves was luteolin-7-O-hexoside-glucuronide (5.84 mg/g DW), whereas the dominant root component was chrysoeriol-7-O-hexoside-glucuronide (0.83 mg/g DW). Infusions from leaves, roots, and their 1:1 (v/v) mixture contained potentially bioaccessible antiradical compounds. S. aloides is a good source of water-extractable reductive compounds. Especially valuable are the leaves of this plant. The roots of S. aloides contained very active hydrophilic compounds able to chelate metal ions. However, their potential bioaccessibility was relatively low. The hydrophilic compounds from the leaves were the most effective XO inhibitors (EC50 = 9.91 mg DW/mL). The water-extractable compounds derived from the leaves and roots acted as uncompetitive LOX inhibitors.

Chemical Characteristics and Anticancer Activity of Essential Oil from Arnica Montana L. Rhizomes and Roots.

Arnica montana L. is a medicinal plant with diverse biological activities commonly used in pharmacy and cosmetics. The attributes of A. montana are mainly related to the concentration and chemical composition of essential oils (EOs). Therefore, the objective of this study was to characterize the chemical composition of EOs derived from A. montana rhizomes and roots taking into account the age of the plants and to investigate the effect of the analyzed EOs on induction of apoptosis, necrosis, and autophagy in human glioblastoma multiforme T98G and anaplastic astrocytoma MOGGCCM cell lines. Rhizomes and roots of mountain arnica were harvested at the end of the third and fourth vegetation periods. The chemical composition of essential oils was determined with the GC-MS technique. Among the 37 components of the essential oil of A. montana, 2,5-dimethoxy-p-cymene (46.47%-60.31%), 2,6-diisopropylanisole (14.48%-23.10%), thymol methyl ether (5.31%-17.79%), p-methoxyheptanophenone (5.07%-9.65%), and α-isocomene (0.68%-2.87%), were detected in the rhizomes and roots of the three-year-old plants and in the rhizomes and roots of the four-year-old plants. The plant part (rhizome, root) and plant age can be determinants of the essential oil composition and, consequently, their biological activity. The induction of apoptosis (but not autophagy nor necrosis) at a level of 28.5%-32.3% is a promising result, for which 2,5-dimethoxy-p-cymene, 2,6-diisopropylanisole, thymol methyl ether, and p-methoxyheptanophenone are probably mainly responsible. The present study is the first report on the anticancer activities of essential oils from A. montana rhizomes and roots.

Chemical Composition of Essential Oil from Flower Heads of Arnica Chamissonis Less. under a Nitrogen Impact.

Chamisso arnica (Arnica chamissonis Less.) is a valuable plant species used in the pharmaceutical industry due to the content of many pharmacologically active substances and the similarity of its chemical composition to that of Arnica montana-a medicinal plant commonly used in pharmacy and cosmetics. The similarity of the two plant species implies that chamisso arnica can be a pharmaceutical substitute for the mountain arnica, i.e., an endangered and endemic plant species in Europe. Chamisso arnica extracts exhibit anti-inflammatory and antiradical activity and possesses high antioxidant properties that might be helpful in preventing or delaying the progress of free radical dependent diseases. The attributes of A. chamissonis are mainly related to the content and chemical composition of essential oil. Therefore, the objective of this study was to characterize the chemical composition of essential oil derived from A. chamissonis flower heads under a nitrogen impact. The experiment was performed on experimental fields in mid-eastern Poland on two soil types (sandy and loamy soils). The nitrogen fertilizer was applied as ammonium sulfate (control, 30, 60, 90, and 120 kg N ha-1). Collection of flower heads was carried out in the full flowering phase, which was characterized by the highest content of essential oil. The chemical composition of essential oil was examined using GC-MS. Among the 75 ingredients of the volatile oil of chamisso arnica flower heads, alpha-pinene, cumene, p-cymene, germacrene D, spathulenol, decanal, caryophyllene oxide, beta-pinene, and benzene acetaldehyde were present at relatively high levels. Both the nitrogen application and the soil type had an effect on the oil concentration and the yield of the main constituents (alpha-pinene and germacrene D) with pharmacological value. Different levels of nitrogen application could be considered as a relevant way to modify the chemical composition and to increase the essential oil production.

Essential Oil from Arnica Montana L. Achenes: Chemical Characteristics and Anticancer Activity.

Mountain arnica Arnica montana L. is a source of several metabolite classes with diverse biological activities. The chemical composition of essential oil and its major volatile components in arnica may vary depending on the geographical region, environmental factors, and plant organ. The objective of this study was to characterize the chemical composition of essential oil derived from A. montana achenes and to investigate its effect on induction of apoptosis and autophagy in human anaplastic astrocytoma MOGGCCM and glioblastoma multiforme T98G cell lines. The chemical composition of essential oil extracted from the achenes was examined with the use of Gas Chromatography-Mass Spectrometry GC-MS. Only 16 components of the essential oil obtained from the achenes of 3-year-old plants and 18 components in the essential oil obtained from the achenes of 4-year-old plants constituted ca. 94.14% and 96.38% of the total EO content, respectively. The main components in the EO from the arnica achenes were 2,5-dimethoxy-p-cymene (39.54 and 44.65%), cumene (13.24 and 10.71%), thymol methyl ether (8.66 and 8.63%), 2,6-diisopropylanisole (8.55 and 8.41%), decanal (7.31 and 6.28%), and 1,2,2,3-tetramethylcyclopent-3-enol (4.33 and 2.94%) in the 3- and 4-year-old plants, respectively. The essential oils were found to exert an anticancer effect by induction of cell death in anaplastic astrocytoma and glioblastoma multiforme cells. The induction of apoptosis at a level of 25.7-32.7% facilitates the use of this secondary metabolite in further studies focused on the development of glioma therapy in the future. Probably, this component plays a key role in the anticancer activity against the MOGGCCM and T98G cell lines. The present study is the first report on the composition and anticancer activities of essential oil from A. montana achenes, and further studies are required to explore its potential for future medicinal purposes.

Long term insight into biodiversity of a smelter wasteland reclaimed with biosolids and by-product lime.

Smelter wastelands containing high amounts of zinc, lead, cadmium, and arsenic constitute a major problem worldwide. Serious hazards for human health and ecosystem functioning are related to a lack of vegetative cover, causing fugitive dust fluxes, runoff and leaching of metals, affecting post-industrial ecosystems, often in heavily populated areas. Previous studies demonstrated the short term effectiveness of assisted phytostabilisation of zinc and lead smelter slags, using biosolids and liming. However, a long term persistence of plant communities introduced for remediation and risk reduction has not been adequately evaluated. The work was aimed at characterising trace element solubility, plant and microbial communities of the top layer of the reclaimed zinc and lead smelter waste heaps in Piekary Slaskie, Poland, 20 years after the treatment and revegetation. The surface layer of the waste heaps treated with various rates of biosolids and the by-product lime was sampled for measuring chemical and biochemical parameters, which are indicative for metals bioavailability as well as for microorganisms activity. Microbial processes were characterised by enzyme activities, abundance of specific groups of microorganisms and identification of N fixing bacteria. Plant communities of the area were characterised by a percent coverage of the surface and by a composition of plant species and plant diversity. The study provides a strong evidence that the implemented remediation approach enables a sustainable functioning of the ecosystem established on the toxic waste heaps. Enzyme activities and the count of various groups of microorganisms were the highest in areas treated with both biosolids and lime, regardless their rates. A high plant species diversity and microbial activities are sustainable after almost two decades from the treatment, which is indicative of a strong resistance of the established ecosystem to a metal stress and a poor physical quality of the anthropogenic soil formed by the treatment.

Metal accumulation strategies in plants spontaneously inhabiting Zn-Pb waste deposits.

Metal (Zn, Pb, Cd, Cu, Ni, Cr) accumulation in shoots of 38 plant species spontaneously colonizing three Zn-Pb waste deposits in southern Poland was studied in order to find out if the age of the waste (30-130 years) or its type (slag or flotation residues) influence metal content in plants and to identify species potentially suitable for biomonitoring and phytoremediation. The total metal concentrations in the waste upper layers ranged from 7300 to 171,790 mg kg(-1) for Zn, from 1390 to 22,265 mg kg(-1) for Pb, and from 66 to 1,464 mg kg(-1) for Cd, whereas CaCl2-extracted fractions accounted for 0.034-0.11 %, 0.005-0.03 %, and 0.28-0.62 % of total Zn, Pb and Cd concentrations, respectively. The concentrations of Cu, Ni, and Cr in substrates and in plants were low and ranged within the background values. Metal accumulation in plant shoots was poorly correlated with both total and CaCl2-extracted forms of metals in the substrate and was highly variable among species and also specimens of the same species. The highest mean concentrations of Zn, Pb and Cd were found in Anthyllis vulneraria L. (901.5 mg kg(-1)), Echium vulgare L. (116.92 mg kg(-1)), and Hieracium piloselloides Vill. (26.86 mg kg(-1)), respectively. Besides Reseda lutea L., no species appeared to be a good indicator of polymetallic environment pollution based on chemical analysis of shoots; however, metal accumulation in the whole plant communities of a particular contaminated area might be an accurate tool for assessment of metal transfer to vegetation irrespective of the type or age of the waste. All the species studied developed a metal exclusion strategy, thus exhibiting potential for phytostabilization of metalliferous wastelands.

Bioaccessibility in vitro of nutraceuticals from bark of selected Salix species.

The aim of this study was to investigate and to compare the extractability, bioaccessibility, and bioavailability in vitro of antioxidative compounds from bark of selected Salix species: S. alba (SA), S. daphnoides (SD), S. purpurea (SP), and S. daphnoides x purpurea (SDP) hybrid willow clones originating from their natural habitats and cultivated on the sandy soil. The highest amount of phenolic glycosides was found in the bark of SDP and SD. The best source of phenolics was bark of SDP. The highest content of flavonoids were found in SD bark samples, whereas the highest concentration of bioaccessible and bioavailable phenolic acids was determined in SDP bark. Bark of all tested Salix species showed significant antiradical activity. This properties are strongly dependent on extraction system and genetic factors. Regardless of Salix genotypes, the lowest chelating power was found for chemically-extractable compounds. Bark of all Salix species contained ethanol-extractable compounds with reducing ability. Besides this, high bioaccessibility and bioavailability in vitro of Salix bark phytochemicals were found. Obtained results indicate that extracts from bark tested Salix genotypes can provide health promoting benefits to the consumers; however, this problem requires further study.

Propagation and introduction of Arnica montana L. into cultivation: a step to reduce the pressure on endangered and high-valued medicinal plant species.

Arnica montana (L.) is an endangered and endemic medicinal plant species in Europe. The pressure on natural sources of this plant is alleviated by a suitable use of arnica resources in the European region and introduction into cultivation. The objective of this study was to describe the impact of different ways of plant propagation and introduction on the growth and reproduction mode of this species. During the six consecutive years of the field experiment, the vegetative and reproductive traits were monitored, and survival time was assessed. The particular ways of arnica plant propagation and introduction determined all the intrinsic species traits and plant survival. The values of the characteristics studied indicated good acclimatization of the arnica ecotype to the climatic conditions of eastern Poland. Practical implications from the data presented here include the possibility of using the presented modes of arnica propagation and introduction in the short- and long-term perspective of arnica cultivation, which can give a possibility of better adjustment of raw material production.