Ligninolytic fungi in bioremediation: extracellular enzyme production and degradation rate

Ligninolytic fungi can be used for remediation of pollutants in water and soil. Extracellular peroxidases and laccases have been shown to oxidize recalcitrant compounds in vitro but the likely significance of individual enzyme levels in vivo remains unclear. This study documents the amounts and activities of Mn-dependent peroxidase (MnP), lignin peroxidase and laccase (LAC) in various species of ligninolytic fungi grown in liquid medium and soil and their effect on degradation of polycyclic aromatic hydrocarbons (anthracene and pyrene), a polychlorinated biphenyl mixture (Delor 106) and a number of synthetic dyes. Stationary cultures of a highly degradative strain Irpex lacteus exhibited 380-fold and 2-fold increase in production of MnP and LAC, respectively, compared to submerged cultures. Addition of Tween 80 to the submerged culture increased MnP levels 260-fold. High levels of MnP correlated with efficient decolorization of Reactive Orange 16 azo dye but not of Remazol Brilliant Blue R anthraquinone dye. Degradation of anthracene and pyrene in spiked soil by straw-grown explorative mycelium of Phanerochaete chrysosporium, Trametes versicolor and Pleurotus ostreatus showed the importance of MnP and LAC levels secreted into the soil. The importance of high fungal enzyme levels for efficient degradation of recalcitrant compounds was better demonstrated in liquid media compared to the same strains growing in soil.     

New view on the age-specificity of pig Cryptosporidium by species-specific primers for distinguishing Cryptosporidium suis and Cryptosporidium pig genotype II

Two species of Cryptosporidium are commonly identified in pigs: Cryptosporidium suis and Cryptosporidium pig genotype II. Detection of Cryptosporidium spp. is routinely based on molecular methods such as polymerase chain reaction (PCR) and subsequent restriction fragment length polymorphism (RFLP) or gene sequencing. However, most of these methods are hampered by low sensitivity to mixed infections. As a solution of this problem, novel species-specific primers were designed and tested in the present study. Sensitivity of our primers was identical to genus-specific primers, but more (1:48) mixed infections were detected using these species-specific primers on 477 DNA samples originating from naturally infected pigs of different age categories. Our results show differences in age-dependent susceptibility of pigs to the infection. Whereas C. suis was found in all tested categories of pigs (1-12 week of age and sows), Cryptosporidium pig genotype II was recorded only in animals older than 6 week of age. Usage of species-specific primers could help to better the understanding of epidemiology of pig specific Cryptosporidium spp. and its occurrence, which, on the basis of our results, is underestimated.     

Artemisia dracunculus L. (tarragon): a critical review of its traditional use, chemical composition, pharmacology, and safety

Artemisia dracunculus L. (tarragon) has a long history of use as a spice and remedy. Two well-described "cultivars" (Russian and French) are used widely and differ in ploidy level, morphology, and chemistry. Key biologically active secondary metabolites are essential oils (0.15-3.1%), coumarins (>1%), flavonoids, and phenolcarbonic acids. In vivo studies mainly in rodents, particularly from Russian sources, highlight potential anti-inflammatory, hepatoprotective, and antihyperglycemic effects. Despite concerns about the toxic effects of two of its main constituents, estragole (up to 82%) and methyleugenol (up to 39%), no acute toxicity or mutagenic activity has been reported at doses relevant for human consumption. Water extracts of A. dracunculus contain very low amounts of estragole and methyleugenol and, therefore, are considered to pose a very limited risk. Overall, a stronger focus on clinical studies and precise taxonomic and phytochemical definition of the source material will be essential for future research efforts.     

Sap flux – a real time assessment of health status in Norway spruce

Top dieback of Norway spruce (Picea abies), triggered by drought in 2004–2006, has been observed in Southeast Norway and trees died within four years after appearance of the first symptoms. The aim of our study was to use sap flux measurements as a diagnostic method for assessment of tree vitality. We used the heat field deformation method to monitor the sap flux density (SFD) in four pairs of healthy and declining trees in situ. To provide retrospective information on hydraulic performance of the trees we took samples for wood anatomical analysis. After felling the trees we used the modified differential translucence method (MDT) as a proxy for the SFD measurements. Healthy trees had three times higher SFD values as declining trees. In some healthy trees we detected decreasing SFD with time. The MDT method agreed with the SFD measurements. In conclusion, we detected sap flux dysfunction in declining trees and showed that the SFD reduction may occur during a short period, prior to occurrence of any visual symptoms. We suggest incorporating the SFD measurements into the repertoire of diagnostic tools in forest pathology.     

Effects of Endo- and Ectomycorrhizal Fungi on Physiological Parameters and Heavy Metals Accumulation of Two Species from the Family Salicaceae

There is increasing interest in poplars and willows due to their biomass production and phytoremediation potential. They host two major types of mycorrhizal fungi that can substantially modulate the physiology of their hosts. In this study, the effects of endo- and ectomycorrhizal fungi on growth, physiological parameters, and heavy metals accumulation were studied in a pot experiment using Salix alba L. and Populus nigra L. The mycorrhizal fungi were inoculated separately and in combination to a soil substrate polluted by a mixture of heavy metals (mainly Cd, Pb, and Zn). Tree species differed in their mycorrhizal affinities, with poplar being colonized predominantly by Glomus intraradices and willow by Hebeloma mesophaeum. H. mesophaeum increased willow height and biomass, while G. intraradices decreased poplar height. The photosynthetic rate remained unchanged, and only minor changes were observed in the relative composition of photosynthetic pigments. Poplar photosynthetic rates and levels of photosynthetic pigments declined, while the epicuticular waxes in leaves increased toward the end of the experiment, irrespective of the inoculation. H. mesophaeum strongly reduced the accumulation of Cd and Fe in willow and poplar shoots, respectively. Our results support the use of selected mycorrhizal strains to tune phytoremediation outcomes in their plant hosts.     

Characterisation of Czech arable weed communities according to management and production area considering the prevalence of herbicide-resistant species

Herbicide resistance is a widespread issue that impacts management of conventional farms, but also has ramifications for the weed community assembly; it is therefore important to see how these species factor into the weed community assembly of farms throughout the countryside. This research analysed species richness and community diversity in 98 field relevés from 48 organic and 50 conventional farms evenly distributed between two major production regions of the Czech Republic and then evaluated the incidence of species which have been reported resistant in the Czech Republic and its neighbouring countries. Farms were selected independently of any acknowledged resistant species. Out of 164 species found in this survey, only eight species have had herbicide-resistant biotypes reported in the Czech Republic, while a total of 19 species had herbicide-resistant biotypes reported in neighbouring countries. Species with recorded resistance to PSII inhibitors in the Czech Republic tended to be found together and were mostly associated with the beet production region, characterised by low altitude: Amaranthus retroflexus, Chenopodium album, Echinochloa crus-galli and Solanum nigrum. Species with reported resistance to ALS and ACCase-inhibiting herbicides were not clearly associated with a particular region or farming type. Of the species which have had reported herbicide resistance in the neighbouring countries, several were found in conventional fields within the growing season and we recommend immediate screening for herbicide resistance in these species and more diligent action in management according to anti-resistance strategies: Bromus sterilis resistant to ACCase-inhibiting herbicides, A. retroflexus resistant to ALS-inhibiting herbicides or Avena fatua resistant to ACCase and ALS-inhibiting herbicides. This work is unique in that it is evaluating weed species diversity in organic and conventional farms and using the context to frame the prevalence of high-risk herbicide-resistant species; thereby putting the potential incidence of herbicide resistance into perspective at the landscape level.     

Platelet-Rich Plasma in the Treatment of Induced Tendinopathy in Horses: Histologic Evaluation

This study was carried out to evaluate the effect of platelet-rich plasma (PRP) on the treatment of tendinopathy induced in the superficial digital flexor tendon (TFDS) of horses, by using histologic evaluation. Six healthy crossbred geldings aged 8 to 15 years (12 ± 3) were used. The TFDS tendinopathy was provoked in both forelimbs, by intratendinous administration of 2.5 mg collagenase (2.5 mg/mL), and this procedure was considered as the beginning of the experimental phase. At 12 days after induction of the tendinopathy, the animals were subjected to the following treatments: (1) in the lesion caused in the right superficial digital flexor tendon (PRP-treated group), 2.5 mL PRP activated with calcium chloride at 0.0125 mol/L at concentrations from 320,000 to 500,000 platelets/μL, were injected; (2) in the tendinopathy of the left SDFT (control group), 2.5 mL 0.9% saline solution was administrated. Thirty-six days after the treatments, a biopsy of the injured area was performed for histologic evaluation. In both groups, the histologic analysis showed an increase in the fibroblastic density, as well as the presence of neovascularization, lymphocytes, and plasmocytes infiltrate and tissue organization at variable intensity. In the PRP-treated group, the SDFT was more organized, with the collagen fibers and fibroblasts being better arranged on the tendon matrix. The numbers of the fibroblasts and blood vessels did not differ between the groups. Histologic evaluation 36 days after tendinopathy showed that injuries under a single PRP treatment present a more uniform and organized tissue repair when compared with the control group.