Radial stem growth in response to microclimate and soil moisture in a drought-prone mixed coniferous forest at an inner Alpine site

Dendroclimatological studies in a dry inner Alpine environment (750 m a.s.l.) revealed different growth response of co-occurring coniferous species to climate, which is assumed to be caused by a temporal shift in wood formation among species. The main focus of this study therefore was to monitor intra-annual dynamics of radial increment growth of mature deciduous and evergreen coniferous species (Pinus sylvestris, Larix decidua and Picea abies) during two consecutive years with contrasting climatic conditions. Radial stem growth was continuously followed by band dendrometers and modelled using Gompertz functions to determine time of maximum growth. Histological analyses of tree ring formation allowed determination of temporal dynamics of cambial activity and xylem cell development. Daily fluctuations in stem radius and radial stem increments were extracted from dendrometer traces, and correlations with environmental variables were performed. While a shift in temporal dynamics of radial growth onset and cessation was detected among co-occurring species, intra-annual radial growth peaked synchronously in late May 2011 and early June 2012. Moist atmospheric conditions, i.e. high relative air humidity, low vapour pressure deficit and low air temperature during the main growing period, favoured radial stem increment of all species. Soil water content and soil temperature were not significantly related to radial growth. Although a temporal shift in onset and cessation of wood formation was detected among species, synchronous culmination of radial growth indicates homogenous exogenous and/or endogenous control. The close coupling of radial growth to atmospheric conditions points to the importance of stem water status for intra-annual growth of drought-prone conifers.     

Status of aquatic and riparian biodiversity in artificial lake ecosystems with and without management for recreational fisheries: Implications for conservation

1. Humanity is facing a biodiversity crisis, with freshwater-associated biodiversity in a particularly dire state. Novel ecosystems created through human use of mineral resources, such as gravel pit lakes, can provide substitute habitats for the conservation of freshwater and riparian biodiversity. Many of these artificial ecosystems are subject to a high intensity of recreational use, however, which may limit their biodiversity potential.
2. The species richness of several taxa (plants, amphibians, dragonflies, damselflies, waterfowl, and songbirds) was assessed and a range of taxonomic biodiversity metrics were compared between gravel pit lakes managed for recreational fisheries (n = 16) and unmanaged reference lakes (n = 10), controlling for non-fishing-related environmental variation.
3. The average species richness of all the taxa examined was similar among lakes in both lake types and no substantial differences in species composition were found when examining the pooled species inventory. Similarly, there were no differences between lake types in the presence of rare species and in the Simpson diversity index across all of the taxa assessed.
4. Variation in species richness among lakes was correlated with woody habitat, lake morphology (surface area and steepness), and land use, but was not correlated with the presence of recreational fisheries. Thus, non-fishing-related environmental variables had stronger effects on local species presence than recreational fisheries management or the presence of recreational anglers.
5. Collectively, no evidence was found that anglers and recreational fisheries management constrain the development of aquatic and riparian biodiversity in gravel pit lakes in the study region; however, the conservation of species diversity in gravel pit lakes could benefit from an increasing reliance on habitat enhancement activities.

     

Accurate Dereplication of Bioactive Secondary Metabolites from Marine-Derived Fungi by UHPLC-DAD-QTOFMS and a MS/HRMS Library

In drug discovery, reliable and fast dereplication of known compounds is essential for identification of novel bioactive compounds. Here, we show an integrated approach using ultra-high performance liquid chromatography-diode array detection-quadrupole time of flight mass spectrometry (UHPLC-DAD-QTOFMS) providing both accurate mass full-scan mass spectrometry (MS) and tandem high resolution MS (MS/HRMS) data. The methodology was demonstrated on compounds from bioactive marine-derived strains of Aspergillus, Penicillium, and Emericellopsis, including small polyketides, non-ribosomal peptides, terpenes, and meroterpenoids. The MS/HRMS data were then searched against an in-house MS/HRMS library of ~1300 compounds for unambiguous identification. The full scan MS data was used for dereplication of compounds not in the MS/HRMS library, combined with ultraviolet/visual (UV/Vis) and MS/HRMS data for faster exclusion of database search results. This led to the identification of four novel isomers of the known anticancer compound, asperphenamate. Except for very low intensity peaks, no false negatives were found using the MS/HRMS approach, which proved to be robust against poor data quality caused by system overload or loss of lock-mass. Only for small polyketides, like patulin, were both retention time and UV/Vis spectra necessary for unambiguous identification. For the ophiobolin family with many structurally similar analogues partly co-eluting, the peaks could be assigned correctly by combining MS/HRMS data and m/z of the [M + Na]⁺ ions.     

Acoustic emission analysis of industrial plywood materials exposed to destructive tensile load

Several plywood materials made from spruce wood and, for comparison, solid spruce wood were investigated focusing on the sub-macroscopic damage evolution during tensile loading of the specimens. The destructive tests were simultaneously monitored by the acoustic emission (AE) method and strain field deformation measurement using digital image correlation (DIC). The bilinear interpretation of exponential defect growth identified the start of significant nonlinear behavior at 70 % of ultimate strength for all plywood materials. However, already the preceding and more stable damage evolution at lower stress levels has indicated a variation in intensity of the source mechanisms evaluated by AE energy of the detected events. Additional information on the formation of strain field concentration, which correlates with discrete accumulation in AE events and increased spreading in the distribution of AE energy, reveals the complexity of pre-damage due to the variation in cracks’ magnitude and timescales involved. The correlation between ultimate tensile strength and damage accumulation below 70 % of ultimate strength is determined, as well as the influence of layered structures on damage size shown by the percentage distribution of AE energy.     

The effects of light and temperature on the photosynthesis of the Asparagopsis armata tetrasporophyte (Falkenbergia rufolanosa), cultivated in tanks

The integrated aquaculture of the tetrasporophyte of Asparagopsis armata Harvey (Falkenbergia rufolanosa) using fish farm effluents may be viable due to the species high capacity of removing nutrients and its content of halogenated organic compounds with applications on the pharmaceutical and chemical industries. In order to optimize the integrated aquaculture of F. rufolanosa, we followed the daily variation of the potential quantum yield (F_v/F_m) of PSII on plants cultivated at different biomass densities and different total ammonia nitrogen (TAN) fluxes to check if they are photoinhibited at any time of the day. Moreover, the photoinhibition under continuous exposure to highly saturating irradiance and its potential for subsequent recovery in the shade was assessed. The potential for year round cultivation was evaluated by measuring rates of O₂ evolution of plants acclimated at temperatures ranging from 15 to 29 °C, the temperature range of a fish farm effluent in southern Portugal where an integrated aquaculture system of F. rufolanosa was constructed.Photoinhibition does not seem to be a major constrain for the integrated aquaculture of F. rufolanosa. Only when cultivated at a very low density of 1.5 g fresh weight (FW) l^− 1 that there was a midday decrease in maximal quantum yield (F_v/F_m). At densities higher than 4 g FW l^− 1, no photoinhibition was observed. When exposed to full solar irradiance for 1 h, F. rufolanosa showed a 33% decrease in F_v/F_m, recovering to 86% of the initial value after 2 h in the shade. A midday decline of the F. rufolanosa F_v/F_m was also observed under the lowest TAN flux tested (∼6 μM h^− 1), suggesting that this fast and easy measurement of fluorescence may be used as a convenient diagnostic tool to detect nutrient-starved unbalance conditions of the cultures. Maximum net photosynthesis peaked at 15 °C with 9.7 mg O₂ g dry weight (DW)^− 1 h^− 1 and remained high until 24 °C. At 29 °C, the net oxygen production was significantly reduced due to a dramatic increase of respiration, suggesting this to be the species' lethal temperature threshold.Results indicate that F. rufolanosa has a considerable photosynthetic plasticity and confirm it as a good candidate for integrated aquaculture at temperatures up to 24° C and cultivation densities of at least 5 g FW l^− 1. When cultivated at these densities, light does not penetrate below the first few centimetres of the surface zone. Plants circulate within the tanks, spending around 10% of the time in the first few centimetres where they are able to use efficiently the saturating light levels without damaging their photosynthetic apparatus.     

Bitter-tasting and kokumi-enhancing molecules in thermally processed avocado (Persea americana Mill.)

Sequential application of solvent extraction and RP-HPLC in combination with taste dilution analyses (TDA) and comparative TDA, followed by LC-MS and 1D/2D NMR experiments, led to the discovery of 10 C(17)-C(21) oxylipins with 1,2,4-trihydroxy-, 1-acetoxy-2,4-dihydroxy-, and 1-acetoxy-2-hydroxy-4-oxo motifs, respectively, besides 1-O-stearoyl-glycerol and 1-O-linoleoyl-glycerol as bitter-tasting compounds in thermally processed avocado (Persea americana Mill.). On the basis of quantitative data, dose-over-threshold (DoT) factors, and taste re-engineering experiments, these phytochemicals, among which 1-acetoxy-2-hydroxy-4-oxo-octadeca-12-ene was found with the highest taste impact, were confirmed to be the key contributors to the bitter off-taste developed upon thermal processing of avocado. For the first time, those C(17)-C(21) oxylipins exhibiting a 1-acetoxy-2,4-dihydroxy- and a 1-acetoxy-2-hydroxy-4-oxo motif, respectively, were discovered to induce a mouthfulness (kokumi)-enhancing activity in sub-bitter threshold concentrations.     

Negligible contribution from roots to soil-borne phospholipid fatty acid fungal biomarkers 18:2ω6,9 and 18:1ω9

The phospholipid fatty acid biomarkers 18:1ω9, 18:2ω6,9 and 18:3ω3,6,9 are commonly used as fungal biomarkers in soils. They have, however, also been found to occur in plant tissues, such as roots. Thus, the use of these PLFAs as fungal biomarkers in sieved soil, which may still contain small remains of roots, has been questioned. We used data from a recent beech tree girdling experiment to calculate the contribution of roots to these biomarkers and were able to demonstrate that not more than 0.61% of 18:1ω9 and 18:2ω6,9 in sieved soil samples originated from roots (but 4% of 18:3ω3,6,9). Additionally, the abundance of the biomarker 18:2ω6,9 in the soil was found to be highly correlated to ectomycorrhizal root colonization, which further corroborates its fungal origin. PLFA biomarkers were substantially reduced in vital roots from girdled trees compared to roots of control trees (by up to 76%), indicating that the major part of PLFAs measured in roots may actually originate from ectomycorrhizal fungi growing inside the roots. We calculated, that even a near to 50% reduction in fine root biomass - as observed in the girdling treatment - accounted for only 0.8% of the measured decrease of 18:2ω6,9. Our results demonstrate that both 18:1ω9 and 18:2ω6,9 are suitable biomarkers for detecting fungal dynamics in soils and that especially 18:2ω6,9 is a reliable biomarker to study mycorrhizal dynamics in beech forests.     

The persistence of bovine enterovirus and pseudorabies virus in liquid cattle manure at different storage temperatures

The tenacity of viruses in liquid manure of cattle was examined in a total of five samples inoculated with ECBO-virus (strain LCR-4) representing viruses without envelope and Aujeszky virus (field isolate) representing enveloped viruses. The titers were examined at regular intervals over a period of 26 weeks. On the day of inoculation each sample had a titer of 10(5) ID50/ml. After 16 weeks complete inactivation was observed in the Aujeszky virus sample stored at 20 degrees C. The Aujeszky virus sample wich was kept at 4 degrees C at 26 weeks had a titer of 10(1,75) ID50/ml. In the samples inoculated with ECBO virus after 26 weeks of inoculation a titer of 10(3) ID50/ml was found in the manure stored at 20 degrees C. No influence on the virus titers in the liquid manure samples was observed either from pH or the number of bacteria (3,4 x 10(7)-1.16 x 10(8)/ml during the examination period.     

Establishment of proliferative cell nuclear antigen gene as an internal reference gene for polymerase chain reaction of a wide range of archival and fresh mammalian tissues.

Polymerase chain reaction (PCR) from paraffin-embedded tissues provides a powerful tool to amplify DNA from a variety of recent and archival material. Because DNA from paraffin-embedded samples is more degraded than from fresh material, the amplification of reference genes is essential to exclude false-negative results. This study describes the use of the proliferative cell nuclear antigen (PCNA) gene as a reference gene in a range of animal species and in humans. The PCNA-PCR to amplify a fragment extending from exon 5 through exon 6 and including the intervening intron 6 gave a reproducible pattern, with a 280-base pair (bp) band from canine, equine, bovine, ovine, and caprine samples showing high sequence homology. Porcine, guinea pig, tiger, and lion samples, however, gave an additional fragment of approximately 197 bp. The whole intron 6 from these fragments is missing, possibly representing a pseudogene. In feline samples only the 197-bp fragment could be detected. This study shows that the PCNA gene is highly conserved across a broad range of animal species and is well suited as an internal control for PCR analysis in veterinary medicine.     

Transformation of organoarsenical species by the microflora of freshwater crayfish

A study of the transformation of arsenic species by the microflora of the freshwater crayfish Procambarus clarkii was carried out. The study of the degradation of AB (arsenobetaine) was performed in aerobic conditions in two culture media (tryptic soy broth and saline medium) at two temperatures (30 and 8 degrees C). The microflora transformed AB into TMAO (trimethylarsine oxide), DMA (dimethylarsinate), MA (methylarsonate), and an unidentified compound (U1). The quickest transformations were carried out by microflora from hepatopancreas incubated in saline medium at 30 degrees C. The individualized study of other arsenic species [AC (arsenocholine), TETRA (tetramethylarsonium ion), TMAO, DMA, and MA] was also performed in saline medium. The only transformation observed was of AC into AB. The bacteria possibly responsible for AB degradation were isolated, identified by phenotypic and genotypic methods, and individually assayed for AB transformation. Only isolates allocated to the species Pseudomonas putida were able to metabolize AB.