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The applicability of herbicidal ionic liquids (HILs) as an alternative form of herbicides is currently evaluated. Yet, the available research is lacking information on the behaviour of herbicidal ionic liquids upon addition to the environment, i.e., if cations and anions act as separate moieties or remain an ionic salt. Hence, we tested degradation of five HILs with the glyphosate anion, their bioavailability in soil, toxicity towards microorganisms, impact on the biodiversity and the abundance of phnJ and soxA genes. The cations were proven to be slightly or moderately toxic. The properties of cations determined the properties of the whole formulation, which it might suggest that cations and anion act as the independent mixture of ions. The mineralisation efficiencies were in the range of 15–53%; however, in the case of cations (except non-toxic choline), only 13–20% were bioavailable for degradation. The hydrophobic cations were proven to be highly sorbed, while the anion was readily available for microbial degradation regardless of its counterion. The approach to enrich test samples with isolated microorganisms specialised in glyphosate degradation resulted in higher degradation efficiencies, yet not high enough to mitigate the negative impact of cations. In addition, increased activity of enzymes participating in glyphosate degradation was observed. In the view of obtained results, the use of cationic surfactants in HILs structure is not recommended, as sorption was shown to be determining factor in HILs degradation efficiency. Moreover, obtained results indicate that corresponding ions in HILs might act as separate moieties in the environment.

Ionic liquids (ILs) are described as low-impact green chemicals in which the cation can influence the properties of the anion through ion pair formation. We decided to see how stable ILs are in the soil environment. Herbicidal ILs consisting of hydrophilic glyphosate [Glyph] paired with hydrophilic choline [Chol], hydrophobic C12 alkyl modified choline [C12Chol] and hydrophobic quaternary ammonium cation [C16TMA] were synthesized. The calculated interaction energies between ions (75.9–88.9 kcal/mol) were typical for ILs. The results showed independent sorption of cations and anions and no statistical effect of cations on anion sorption. Hydrophobic [C16TMA], [C12Chol] cations followed 99 % adsorption, and hydrophilic [Chol] revealed 42–45 % adsorption. The Freundlich constants were [C16TMA] Kf≈4000 > [C12Chol] Kf≈3000 > [Chol] Kf≈3 (mg1-1/n L1/n g-1). The preferential sorption of hydrophobic cations was also visible during leaching experiments and resembles the behavior of cationic surfactants. [Glyph] sorption was higher in organic-rich agricultural soil (approx. 45 %) compared to reference OECD soil (30 %), and the Kf values were constant, regardless of the presence of cations. The lack of ionic pair integrity was also visualized by the toxicity and phytotoxicity results. In the soil environment, ILs are a mixture of independent cations and anions rather than a novel emerging contaminant.

Forests accumulate and store large amounts of carbon (C), and a substantial fraction of this stock is contained in deadwood. This transient pool is subject to decomposition by deadwood-associated organisms, and in this process it contributes to CO2 emissions. Although fungi and bacteria are known to colonize deadwood, little is known about the microbial processes that mediate carbon and nitrogen (N) cycling in deadwood. In this study, using a combination of metagenomics, metatranscriptomics, and nutrient flux measurements, we demonstrate that the decomposition of deadwood reflects the complementary roles played by fungi and bacteria. Fungi were found to dominate the decomposition of deadwood and particularly its recalcitrant fractions, while several bacterial taxa participate in N accumulation in deadwood through N fixation, being dependent on fungal activity with respect to deadwood colonization and C supply. Conversely, bacterial N fixation helps to decrease the constraints of deadwood decomposition for fungi. Both the CO2 efflux and N accumulation that are a result of a joint action of deadwood bacteria and fungi may be significant for nutrient cycling at ecosystem levels. Especially in boreal forests with low N stocks, deadwood retention may help to improve the nutritional status and fertility of soils.

Too low concentrations of organic pollutants as well as too high loads of toxic contaminants and/or other extreme conditions of (waste)waters may impede their efficient treatment by conventional biological technologies. Micropollutants typically occur in only minute amounts in the aquatic environment, which is not in favor of their productive biodegradation and may lead to infinitely low degradation rates. Conventional bioprocesses could even collapse in response to too toxic or otherwise extreme conditions of the waters to be treated. Advanced biological water treatment processes aim to overcome the performance limits of conventional biological processes, and make use of isolated enzymes and whole organisms. Current approaches involve enzyme immobilization to yield biocatalytically active nanomaterials, and bioelectrochemical systems employing immobilized microorganisms for wastewater treatment along with energy production. Such applications are mostly still at the experimental stage, and issues related to process upscaling, long-term stability, and cost efficiency still need to be solved. Membrane bioreactors (MBRs) retain the biocatalytically active sludge in the system. They can be augmented with additional degrader organisms or used in combination with physicochemical processes such as activated carbon adsorption and ozonation. MBRs have already successfully been tested at the pilot and sometimes even at full scale. Constructed wetlands also have also the potential to remove micropollutants from urban wastewater. They may be considered as an alternative for wastewater treatment in small and scattered communities, and for the final step in the treatment of special wastewaters.

Due to their enhanced reactivity, metal and metal-oxide nanoscale zero-valent iron (nZVI) nanomaterials have been introduced into remediation practice. To ensure that environmental applications of nanomaterials are safe, their possible toxic effects should be described. However, there is still a lack of suitable toxicity tests that address the specific mode of action of nanoparticles, especially for nZVI. This contribution presents a novel approach for monitoring one of the most discussed adverse effects of nanoparticles, i.e., oxidative stress (OS). We optimized and developed an assay based on headspace-SPME-GC-MS analysis that enables the direct determination of volatile oxidative damage products (aldehydes) of lipids and proteins in microbial cultures after exposure to commercial types of nZVI. The method employs PDMS/DVB SPME fibers and pentafluorobenzyl derivatization, and the protocol was successfully tested using representatives of bacteria, fungi, and algae. Six aldehydes, namely, formaldehyde, acrolein, methional, benzaldehyde, glyoxal, and methylglyoxal, were detected in the cultures, and all of them exhibited dose-dependent sigmoidal responses. The presence of methional, which was detected in all cultures except those including an algal strain, documents that nZVI also caused oxidative damage to proteins in addition to lipids. The most sensitive toward nZVI exposure in terms of aldehyde production was the yeast strain Saccharomyces cerevisiae, which had an EC50 value of 0.08 g/L nZVI. To the best of our knowledge, this paper is the first to document the production of aldehydes resulting from lipids and proteins as a result of OS in microorganisms from different kingdoms after exposure to iron nanoparticles.

Metal-polluted soils represent hostile environments affecting the composition and functions of soil microbial communities. This study evaluated the implication of combining the mycoremediated dry olive residue (MDOR) amendment application with the inoculation of the arbuscular mycorrhizal fungi (AMF) Funneliformis mosseae in restoring the quality, composition, and functionality of soil microbial communities. To achieve this aim, a mesocosms experiment was set up that included three variations: i) with and without application of Penicillium chrysogenum-10-transformed MDOR (MDOR_Pc), and Chondrosterum purpureum-transformed MDOR (MDOR_Cp) amendments; ii) with and without F. mosseae inoculation; and iii) 30-day and 60-day soil treatment time. As a result of this combined treatment, changes in the soil labile organic C and N fractions were observed throughout the experiment. Increases in the abundance of phospholipid fatty acids (PLFAs) for bacteria, actinobacteria, and Gram− and Gram+ bacteria were also recorded at the end of the experiment. The addition of MDOR amendments boosted fungal and AM fungi communities. AM fungi root and soil colonization was also enhanced as the result of improvement nutrient turnover and spatial conditions caused by adding MDOR in combination with an inoculation of F. mosseae. The composition and functionality of microbial communities seemed to be an important ecological attribute indicating an apparently fully functional restoration of this metal-polluted soil and therefore suggesting the suitability of the combined MDOR and AM fungus treatment as a reclamation practice.

A group of white rot fungi (Irpex lacteus, Panus tigrinus, Dichomitus squalens, Trametes versicolor and Pleurotus ostreatus) was investigated for the biodegradation of norfloxacin (NOR), ofloxacin (OF) and ciprofloxacin (CIP). The selected fluoroquinolones were readily degraded almost completely by I. lacteus and T. versicolor within 10 and 14 d of incubation in liquid medium, respectively. The biodegradation products were identified by liquid chromatography–mass spectrometry. The analyses indicated that the fungi use similar mechanisms to degrade structurally related antibiotics. The piperazine ring of the molecules is preferably attacked via either substitution or/and decomposition. In addition to the degradation efficiency, attention was devoted to the residual antibiotic activities estimated using Gram-positive and Gram-negative bacteria. Only I. lacteus was able to remove the antibiotic activity during the course of the degradation of NOR and OF. The product-effect correlations evaluated by Principal Component Analysis (PCA) enabled elucidation of the participation of the individual metabolites in the residual antibacterial activity. Most of the metabolites correlated with the antibacterial activity, explaining the rather high residual activity remaining after the biodegradation. PCA of ligninolytic enzyme activities indicated that manganese peroxidase might participate in the degradation.

Flumequine, a fluoroquinolone antibiotic, is applied preferably in veterinary medicine, for stock breeding and treatment of aquacultures. Formation of drug resistance is a matter of general concern when antibiotics such as flumquine occur in the environment. Thus, biodegradation of flumequine in solution was investigated using five different ligninolytic fungi. Irpex lacteus, Dichomitus squalens, and Trametes versicolor proved most efficient and transformed more than 90% of flumequine within 6 or even 3 days. Panus tigrinus and Pleurotus ostreatus required up to 14 days to remove >90% of flumequine. Analyses of the metabolites by liquid chromatography–mass spectrometry suggest different transformation pathways for the different fungal strains. Structure proposals were elaborated for 8 metabolites. 7-Hydroxy-flumequine and flumequine ethyl ester were identified as common metabolites produced by all ligninolytic fungi. The largest variety of metabolites was formed by D. squalens. Residual antibacterial activity of the metabolite mixtures was tested using gram-positive and gram-negative bacteria. While for the less efficient P. tigrinus and P. ostreatus cultures the antibacterial activities corresponded to the residual concentrations of flumequine, a remarkable antibacterial activity remained in the D. squalens cultures although flumequine was transformed to more than 90%. Obviously, antibacterially active transformation products were formed by this fungal strain.

Lagomorphs (a group that consists of pikas, hares, rabbits and allies) are notable for their conservative morphology retained for most of their over 50 million years evolutionary history. On the other hand, their remarkable morphological uniformity partly stems from a considerable number of homoplasies in cranial and dental structures that hamper phylogenetic analyses. The premolar foramen, an opening in the palate of lagomorphs, has been characterized as an important synapomorphy of one clade, Ochotonidae (pikas). Within Lagomorpha, however, its phylogenetic distribution is much wider, the foramen being present not only in all ochotonids but also in leporids and stem taxa; its morphology and incidence also varies considerably across the order, even intraspecifically. In this study, we provide a broad survey of the taxonomic distribution of the premolar foramen in extant and fossil Lagomorpha and describe in detail the morphological variation of this character within the group. Micro-computed tomography was used to examine the hard palate and infraorbital groove morphology in Poelagus (Leporidae) and Ochotona. Scans revealed the course and contacts of the canal behind the premolar foramen and structural differences between the two crown clades. We propose that the premolar foramen has evolved independently in several lineages of Lagomorpha, and we discuss development and function of this foramen in the lagomorph skull. This study shows the importance of comprehensive studies on phylogenetically informative non-dental characters in Lagomorpha.

The present field study documents substantial changes in the soil microbial community (SMC) and organic matter (SOM) in an upland pasture soil resulting from 10 years of “cattle outdoor over-wintering practice”. Soils from a long-term investigated pasture area were compared under three different levels of cattle impact (SI – severe, MI – moderate, NI – no impact). Extended polar lipids analysis (PLA) confirmed a qualitatively new microbial community profile and a several-fold increase of the microbial biomass in the impacted soils (SI) compared to the control NI soil. The new SMC was derived from cattle intestine microorganisms, typical by increased content of archaeal phospholipid ether lipids and by new fatty acids indicative for bacterial and fungal fecal anaerobes. A quality of the SI-SOM, evaluated by the relative content of the pyrolytic fragments profile was more similar to the cattle excrements than to the MI and NI soils, and an organic carbon content of the SI soil was not more than three times higher in comparison to the control NI soil. The quality and quantity of the SOM as well as the SMC in both, the most impacted SI and the control NI soils, were stable in contrast to the moderately impacted MI soil. During the growing season, the MI soil lost 75% of the Corg and 65% of the soil microbial biomass that had accumulated during winter; its aromatic-rich-SOM showed transformation into SOM, enriched by N, P-organic derivates. This transformation was positively correlated to a significant recovery of the actinobacteria and reduction of anaerobic microorganisms during the vegetation season. Results in this study showed that the stability of the soil microbial changes due to the cattle outdoor over-wintering husbandry depended on the stability of the quantitative and qualitative changes of the SOM. © 2012 Published by Elsevier B.V.

Endocrine-disrupting compounds (EDCs) represent a large group of substances of natural and anthropogenic origin. They are widely distributed in the environment and can pose serious risks to aquatic organisms and to public health. In this study, 4-n-nonylphenol, technical 4-nonylphenol, bisphenol A, 17a-ethinylestradiol, and triclosan were biodegraded by eight ligninolytic fungal strains (Irpex lacteus 617/93, Bjerkandera adusta 606/93, Phanerochaete chrysosporium ME 446, Phanerochaete magnoliae CCBAS 134/I, Pleurotus ostreatus 3004 CCBAS 278, Trametes versicolor 167/93, Pycnoporus cinnabarinus CCBAS 595, Dichomitus squalens CCBAS 750). The results show that under the used conditions the fungi were able to degrade the EDCs within 14 d of cultivation with exception of B. adusta and P. chrysosporium in the case of triclosane and bisphenol A, respectively. I. lacteus and P. ostreatus were found to be most efficient EDC degraders with their degradation efficiency exceeding 90% or 80%, respectively, in 7 d. Both fungi degraded technical 4-nonylphenol, bisphenol-A, and 17a-ethinylestradiol below the detection limit within first 3 d of cultivation. In general, estrogenic activities assayed with a recombinant yeast test decreased with advanced degradation. However, in case of I. lacteus, P. ostreatus, and P. chrysosporium the yeast assay showed a residual estrogenic activity (28-85% of initial) in 17a-ethinylestradiol cultures. Estrogenic activity in B. adusta cultures temporally increased during degradation of technical 4-nonylphenol, suggesting a production of endocrine-active intermediates. Attention was paid also to the effects of EDCs on the ligninolytic enzyme activities of the different fungi strains to evaluate their possible stimulation or suppression of activities during the biodegradation processes.

Cultures of the ligninolytic fungus Irpex lacteus incubated in a nutrient liquid medium degraded more than 70% of the initially applied benz(a]anthracene within 14 days. At the first step of metabolization, benz[a]anthracene was transformed via a typical pathway of ligninolytic fungi to benz[a]anthracene-7,12-dione (BaAQ). The product was further transformed by at least two ways, whereas one is complied with the anthracene metabolic pathway of L lacteus. Benz[a]anthracene-7,12-dione was degraded to 1,2-naphthalenedicarboxylic acid and phthalic acid that was followed with production of 2-hydroxymethyl benzoic acid or monomethyl and dimethylesters of phthalic acid. Another degradation product of BaAQ was identified as 1-tetralone. Its transformation via 1,4-naphthalenedione, 1,4-naphthalenediol and 1,2,3,4-tetrahydro-1-hydroxynaphthalene resulted again in phthalic acid. None of the intermediates were identified as dead-end metabolites. Metabolites produced by ring cleavage of benz[a]anthracene using the ligninolytic fungus are firstly presented in this work.

The white-rot fungus Irpex lacteus has been reported to be an efficient degrader of polycyclic aromatic hydrocarbons, polychlorinated biphenyls and pentachlorophenol. The fungus produces ligninolytic enzymes laccase, lignin peroxidase and manganese peroxidase (MnP), the latter being the major one produced. MnP was purified using anion exchange and size exclusion chromatography. SDS-PAGE showed the purified MnP to be a monomeric protein of 37 kDa (37.5 kDa using MALDI-TOF) with an isoelectric point at 3.55. The pH optimum was relatively broad, from 4.0 to 7.0 with a peak at pH 5.5. Kinetic constants K-m were 8 mu M for H2O2 and 12 or 31 mu M for Mn2+ depending on the substrate. The enzyme did not perform oxidation in the absence of H2O2 or Mn2+. MnP was active at 5-70 degrees C with an optimum between 50-60 degrees C. At temperatures above 65 degrees C the enzyme rapidly lost activity. Degradation of four representatives of PAHs (phenanthrene, anthracene, fluoranthene, and pyrene) was tested and the enzyme showed the ability to degrade them in vitro. Major degradation products of anthracene were identified. The results confirm the role of MnP in PAH degradation by L lacteus, including cleavage of the aromatic ring.

The biodegradation of polychlorinated biphenyls (PCBs), formerly applied in technical mixtures called Delor 103, was realized by the white rot fungus Pleurotus ostreatus. Besides the efficiency of degradation the translocation of PCBs into the fruit bodies grown during cultivation on straw were of special interest. The selectivity of PCBs degradation is discussed on the basis of structural features of the individual congeners. The most important structural factors affecting the degradation efficiency are indicated with molecular descriptors. Basic dependences of the degradation efficiency of congeners on their chlorination grade and ortho chlorosubstitution could be proved. Additionally, other structural parameters such as the number and position of non-substituted carbon in the biphenyl molecule also affect the degradation results. Apart from the removal of Delor 103 the translocation of PCB isomers into the fruit bodies growing up during cultivation was observed. The structural features assisting the transport of PCB congeners from the straw substrate colonized with mushroom mycelium (straw/mycelium) to the mushroom fruit bodies are also relevant for the degradation mechanism but act reversely. Among the chlorination grade, structural parameters such as the number of para and adversely positioned chlorines or the number of occupied para and meta positions contribute to an accumulation of individual PCB congeners in the mushrooms. In total, only few PCB amounts (below 0.1% of the initially present PCBs in straw) were transported into the fruit bodies and a dangerous PCB contamination was never reached while the experiment. A combination of accelerated solvent extraction and GC-MS analysis was the analytical basis for the investigation presented.

Partition of fluorene, phenanthrene and pyrene in the cultures of the basidiomycete Pleurotus ostreatus (oyster mushroom) was determined by GC-MS analysis coupled with SPME directly within the growing mushroom fruit-body. Headspace (HS) SPME of the corresponding straw/mycelium culture provided information on the contents of PAHs remaining, thus enabling the transfer ratios of PAHs to be estimated. ASE-GC-MS was applied by way of comparison.

Representatives of polycyclic aromatic hydrocarbons (PAHs) were degraded by ligninolytic fungus Irpex lacteus. The products were analyzed by GC–Ion trap mass spectrometry. The combination of full scan mass spectra, product ion scans (MS–MS) and derivatization of the degradation products of anthracene, phenanthrene, fluoranthene and pyrene provided further insight in the degradation mechanism initiated by I. lacteus. Particularly, the product ion scans enabled the interpretation of unknown degradation products, even though they were only produced at trace level. Most of the structures suggested were later confirmed with authentic standards.