The Content of Amino Acids in Pasture Vegetation and Their Apparent Digestibility in 2-year-old Horses

Ten samples of plant vegetation and 10 samples of fresh excrement were taken from the same pasture area. The excrement were collected from 10 2-year-old Old Kladruber horses that received the pasture vegetation daily. The apparent digestibility of nitrogen and amino acids in pasture vegetation was determined by using the acid-insoluble ash marker method. In comparison with excrement, the pasture vegetation contained higher levels of Ser, Ala, Leu, and His and higher levels of Pro (P ≤ .01), Met (P ≤ .01), and Arg (P ≤ .05). The mean level of Ile in pasture vegetation was lower than in excrement (P ≤ .05). The apparent digestibility of amino acids from pasture vegetation was high for Pro and Met (86.75 and 89.39%), moderate for Ser, Ala, Leu, His, and Arg (68.61%-76%), and low for Asp, Thr, Glu, Gly, Val, Leu, Tyr, Phe, and Lys (56.15%-66.03%). The digestibility of lysine and Ile was relatively low (56.39% and 56.15%, respectively). The total content of nitrogen per dry matter was 10.98 ± 2.46 g/kg for pasture vegetation and 12.12 ± 2.38 g/kg for excrement, whereas the content of protein nitrogen was 7.20 ± 0.25 g/kg and 6.89 ± 0.21 g/kg of dry matter in pasture vegetation and excrement, respectively. This means that only 65.55% and 56.90% of N is bound in proteins in pasture vegetation and excrement, respectively. Non-protein nitrogen accounts for 34.45% in pasture vegetation and for 43.10% in excrement.     

Predictive Factors of Sensitivity to Elisidepsin,a Novel Kahalalide F-Derived Marine Compound

Elisidepsin (PM02734, Irvalec^®) is a synthetic marine-derived cyclic peptide of the Kahalalide F family currently in phase II clinical development. Elisidepsin was shown to induce rapid oncosis in ErbB3-expressing cells. Other predictive factors of elisidepsin sensitivity remained unknown. A panel of 23 cancer cell lines of different origin was assessed for elisidepsin cytotoxicity and correlated with mutational state, mRNA and protein expression of selected genes. Elisidepsin showed potent and broad cytotoxic effects in our cancer cell line panel, being active at concentrations ranging from 0.4 to 2 μM that may be relevant for clinical settings. We have shown that elisidepsin is more active in cells harboring epithelial phenotype with high E-cadherin and low vimentin expression. In addition, high ErbB3 and Muc1 expression was correlated with sensitivity to elisidepsin, whereas the presence of KRAS activating mutations was associated with resistance. In DU-PM cells with acquired resistance to elisidepsin, ErbB3 expression was decreased, while Bcl2 was increased. DU-PM cells displayed higher sensitivity to ErbB1-inhibitors suggesting possible cross-talk of ErbB1 and ErbB3 signaling pathways. Combinations of elisidepsin with lapatinib and several chemotherapies including 5-FU and oxaliplatin resulted in synergistic effects that offer the potential of clinical use of elisidepsin in combination settings.     

Distribution and Genotypic Characterization of Campylobacter jejuni Isolated from Poultry in Split and Dalmatia County,Croatia

Consumption of poultry contaminated with Campylobacter jejuni has been recognized worldwide as the leading cause of campylobacteriosis. Therefore, the aim of our study was to investigate the prevalence and genotype diversity of Campylobacter jejuni in poultry meat intended for consumption in Split and Dalmatia County, which is the second biggest County in Croatia. Furthermore, we also wanted to discover possibly stable clones of C. jejuni appearing in different samples and periods of time, which would indicate their ability to persist in or adapt to poultry. In the period from March 2008 until June 2010, 834 samples of poultry from various sources were examined using a surface swab technique. Isolation of C. jejuni was performed by Preston broth and Karmali agar. Identification of the isolates was carried out using biochemical tests. C. jejuni was found in 84 of 574 chicken samples (14.6%) and in nine of 260 samples of turkey (3.5%). Pulse‐field gel electrophoresis (PFGE) was used to analyse 61 obtained isolates using SmaI and KpnI. Of 22 different macrorestriction profiles (MRP) that were found, five were detected in poultry from both different locations and periods of time. Samples from 11 locations were found to be contaminated with more than two different genotypes of C. jejuni. Interestingly, the same MRP were found both in poultry declared to be of domestic origin and in the poultry imported from abroad. The prevalence of C. jejuni in poultry samples was in accordance with previously reported results. Genotypic analysis indicated that the population of C. jejuni in Split and Dalmatia County was diverse and that multiple strains of C. jejuni could be found in the same poultry samples. Furthermore, the same genotypes were identified from the samples obtained from different locations and periods of time, which could support the theory of a global existence of certain MRP that are able to persist in or adapt to poultry.     

Pathogenicity of fungi associated with ash dieback symptoms of one‐year‐old Fraxinus excelsior in Montenegro

In addition to Hymenoscyphus fraxineus, two fungi identified as Diaporthe eres aff. and Fusarium sambucinum aff. were also isolated from necrotic bark lesions on declining one‐year‐old Fraxinus excelsior in a forest stand in Montenegro. To examine their involvement in ash decline, a pathogenicity test was performed using under bark inoculations on one‐year‐old Fraxinus excelsior. Hymenoscyphus fraxineus was included as comparison. All three fungal species proved highly pathogenic towards one‐year‐old seedlings although lesion sizes differed significantly between the different species. Hymenoscyphus fraxineus was most aggressive, followed by F. sambucinum aff., while D. eres aff. caused the smallest lesions. This study demonstrates for the first time the ability of isolates in the D. eres and F. sambucinum species complexes to cause decline on one‐year‐old common ash seedlings.     

Machine learning methods for estimation the indicators of phosphogypsum influence in soil

Purpose

The full understanding of the effect of mineral waste-based fertilizer in soil is still unrelieved, because of the extreme complex chemical composition and plethora of their action pathways. The purposes of this paper is to quantify the input of PG into the soil ecosystem process, considering the direct effects of PG as a whole on soil environment using of a plethora of chemical, toxicological, and biological tests.

Materials and methods

Greenhouse experiment includes different PG doses (0, 1%, 3%, 7.5%, 15%, 25%, and 40%) and two-time collection points after treatments—7 and 28 days. For each treatment and each time collection point, we measure (i) soil pH, bioavailable (H₂0 and NH₄COOH-extractable) element content (S, P, K, Na, Mg, Ca, Fe, Zn, Sr, Ba, F); (ii) soil enzyme activities—dehydrogenase, urease, acid phosphatase, FDA; (iii) soil CO₂ respiration activity with and without glucose addition; (iv) Eisenia fetida, Sinapis alba, and Avena sativa responses. Finally, we combine the ordinary chemical, toxicology, and biological measuring of soil properties with state-of-the-art mathematical analysis, namely (i) support vector machines (used for prediction), (ii) mutual information test (variable importance tasks), (iii) t-SNE and LLE algorithms (used for unsupervised classification).

Results and discussion

The results show similarity between the 0%, 1%, and 3% PG treatments in all collection times based on the toxicological and biological properties. Beyond 7.5% PG, some biological test was significantly inhibited in response to trace element stress. Among all tested parameters, soil urease activities, soil respiration activities after glucose addition, S. alba root lengths, and E. fetida survival rates show sensitivity to PG addition. Furthermore, the machine learning algorithms revealed that only several elements (mobile and water-soluble forms of Ca, Ba, Sr, S, and Na; water-soluble F) could be responsible to elevated soil toxicity for those indicators. SVR models were able to predict soil biological and ecotoxicity properties, and increasing numbers of randomly selected training examples from 50 to 90% of initial experimental data significantly improved model performance.

Conclusions

At this study, we demonstrate benefits of unsupervised machine learning methods for investigating toxicity of man-made substances in soil that can be further applied to risk assessments of various toxins, which are of significant interest to environmental protection.

     

Impact of fertilization on cover crops and microbial community on a bauxite‐mined soil undergoing reclamation

The reclamation of bauxite‐mined areas can be favored by the application of organic and/or chemical fertilization to restore the vegetation. Otherwise, the impact of fertilizations on soil microbiota or plant–microbe interactions as land reclamation progresses is less understood. To address this issue, we evaluated the impact of organic and chemical fertilization on plants and soil microbial community within the first 36 months of land reclamation in a bauxite‐mined site. The experiment was set up according to a split‐plot design in which the main plots received fertilizations [non‐fertilized control (NF), chemical fertilization (CF; NPK and rock phosphate), organic fertilization (OF; poultry litter), and CF+OF combined], and the subplots received cover crops [no cover crops (NC), grass (B; Brachiaria), legume (S, Stylosanthes), and B+S combined]. Cover crops biomass yield was assessed annually with five field campaigns per year. We used phospholipid fatty acids (PLFAs) to infer the impacts of mining and restoration practices on actinobacteria, Gram‐negative and Gram‐positive bacteria, arbuscular mycorrhizal fungi, and fungi. Accordingly, PLFAs were determined before bauxite mining (pre‐mining), six months after topsoil reconfiguration (post‐mining), and after 14 and 36 months following the application of the fertilizations and cover crops. PLFAs results indicated that in post‐mining, the living microbiota was significantly lower than in pre‐mining. Cover crops biomass yield was highest for B and B+S fertilized with CF+OF at 14 and 36 months. Both parametric and non‐parametric statistics showed a temporal variation in the response of living microbes to the treatments applied. After 14 months, living microbes showed greatest response to OF, while at 36 months their response was strongest in the treatments with highest plant biomass production (B and B+S). These results suggest that in the early stages of land reclamation, living microbial biomass benefit the most from organic fertilizers. As this initial boost decline, living microbes are more likely to thrive in areas undergoing reclamation where they can develop synergistic interactions with plants.     

Assessment of the retardation of selected herbicides onto Danube sediment based on small column tests

Journal of Soils and Sediments - This study utilizes column tests to investigate the retardation of certain herbicides with different hydrophobicities (atrazine, alachlor and trifluralin) during...     

Contamination history of suspended river sediments accumulated in oxbow lakes over the last 25 years

Background, aims, and scope  Embankment of meandering river systems in many industrial areas results in the formation of artificial oxbow lakes that may act as perennial or intermittent traps for river sediments. Their deposits can be dated using a combination of historical and stratigraphic data, providing a good means to study historical records of contamination transported by rivers. Contamination history over the last few decades is of special significance for Central and Eastern Europe as it can reflect high pollutant levels in the second half of the twentieth century and the subsequent improvement after the fall of the Iron Curtain. The purpose of this study was to investigate recent sediments of an oxbow lake of the Morava River, Czech Republic, their stratigraphic records, sediment architecture, and history of contamination. Materials and methods  Seven ground-penetrating radar (GPR) profiles and three sediment cores up to 4 m deep were studied. The stratigraphy of the cores was inferred from visible-light spectrophotometry, X-ray radiography, grain size analysis, and semiquantitative modal analysis of sandy fractions. The sediments were dated using the ¹³⁷Cs mass activity and combinations of stratigraphic and historical data. The cores were sampled for concentrations of heavy metals and persistent organic pollutants. Wet sampled, lyophilized, and sieved sediments were extracted and analyzed for heavy metals by inductively coupled plasma mass spectrometry (ICP-MS) of aqua regia leachate and for persistent organic pollutants by gas chromatography (GC-ECD and GC-MS). Results  Three distinct sedimentary sequences (S1, S2, and S3) were identified. The basal sequence S1 represents river channel sediments deposited before the formation of the oxbow lake, most likely before the 1930s. The boundary between the S1 and S2 sequence correlates with the level of sediment dredging from 1981 evidenced from historical data. The overlying sequences S2 and S3 represent a postdredging sediment wedge, which progrades into the lake. ¹³⁷Cs dating revealed a distinct Chernobyl 1986 peak at ∼150 cm depth inferring sedimentation rates up to 7.7 cm/year. Sediment contamination abruptly increased from the pre-1930s deposits to the post-1981 deposits. The concentration levels increased two to five times for Pb, As, Zn, and Cu, about 10 to 15 times for Cr, Sb, and Hg, up to 34 times for Cd, and 25 to 67 times for DDTs, PCBs, and PAHs. The concentrations of most contaminants remained approximately constant until the late 1980s when they started to decrease slowly. The decreasing trends were most prominent for heavy metals and anthracene, less prominent for DDTs, and almost absent for PCBs and most PAHs. Discussion  Different temporal and spatial patterns for various contaminants make it possible to distinguish between anthropogenic point sources from local industry (anthracene, Cr, Cd), possible diffuse sources (most PAHs), and geological background (V, Co, Ni, and Mo). The observed recent trends in heavy metal and persistent organic pollutant contamination are generally consistent with data from other Central European rivers. The roughly balanced contamination levels in sediments from the lake and the adjacent river channel suggest that the oxbow lake deposits reflect immediate levels of the contamination bound to suspended particulate matter passing through the river. Conclusions  The investigated oxbow lake accumulated suspended sediment from Morava River, developing a thick sedimentary body. The sediments offer a good time framework to study historical contamination of the river on a decade time scale. Continuous contamination trends can be traced back to the early 1980s. The results show that stratigraphic analysis of cores has a good potential for identification of uninterrupted historical trends and unconformities, e.g., due to dredging. Recommendations and perspectives  Oxbow lakes may provide an alternative to floodplains and reservoir deposits when studying river contamination history. High sedimentation rates of up to 7.7 cm/year offer a very good stratigraphic resolution, making it possible to study contamination patterns on annual or even seasonal time scales. On the other hand, oxbow lakes may represent long-term contamination stores, which are unlikely to be redistributed by river erosion and, hence, may possess significant environmental risks for the farther future.     

Transformation of Petroleum Saturated Hydrocarbons during Soil Bioremediation Experiments

This paper presents transformations of saturated hydrocarbons of petroleum type pollutants during ex situ bioremediation of soil on the pilot heap (halde), during a period of 6 months, within the grounds of Petroleum Refinery Pan?evo (Serbia). Samples for analysis were taken in time intervals of 2 weeks (P₁–P₁₂ samples). Organic substance was extracted by Soxhlet’s method and quantified. Isoprenoid aliphatics, in particular pristane and phytane, and polycyclic aliphatics of sterane and triterpane types in saturated hydrocarbon fractions were analysed by GC-MS (SIM method). Significant amounts of n-alkanes have not been detected. The MS-chromatogram revealed only marginal amounts of pristane and phytane in sample P₁. Pristane and phytane occurred in sample P₈, and in even higher quantities in the final sample P_12. The proceeding bioremediation process was accompanied by the decrease of the relative amounts of pentacyclic terpanes of hopane type, compared to tri- and tetracyclic terpanes. In the initial sample P₁ the distribution of steranes and hopanes follows a pattern, which is characteristic for crude oils. However, their identification by SIM method was not possible in samples P₈ and P₁₂ because of the reduced concentration. The observed changes in the alkane fractions’ compositions may be considered as atypical, referring to the fact that during oil biodegradation under natural conditions, decomposition of isoprenoids occurs much easier and faster than decomposition of polycyclic alkanes of tri-, tetra- and pentacyclic terpane, sterane and diasterane types, after the decomposition of n-alkanes has been almost completed.