Riboflavin content in the coelomocytes of contrasting earthworm species is differentially affected by edaphic variables including organic matter and metal content

The riboflavin content in extruded coelomocyte lysates derived from Dendrodrilus rubidus may serve as a sensitive bioindicator of soil metal pollution: the vitamin (B2) content has previously been found to be high in worms from unpolluted soil but low in worms inhabiting Zn/Pb mine soils, aerially deposited Ni-contaminated soil, and in worms experimentally transferred from clean soil to the metalliferous field soils. The aim of the present work was to extend these observations by comparing the number and riboflavin composition of coelomocytes retrieved from three lumbricid species (Allolobophora chlorotica, Dendrobaena veneta, Eisenia andrei) after 4-week exposures to an unpolluted commercial soil, two geochemically contrasting unpolluted field soils, and two different Zn/Pb/Cd-polluted soils from the Bukowno district in South Poland. Whilst eco-physiologically contrasting, these three earthworm species share the trait of possessing relatively high numbers of eleocytes, a category of immune-competent coelomocyte rich in autofluorescent riboflavin. Spectroflurometric analysis indicated that coelomocyte riboflavin content in worms maintained in strongly metalliferous soils or in unpolluted sandy-clay and loamy-sand soils was increased in coleomocytes from epigeic D. veneta and E. andrei species, whilst was decreased in endogeic A. chlorotica. In conclusion, the riboflavin content of earthworm coelomocytes is affected in species-specific ways by edaphic variables, including organic matter and metal pollution.     

Determination of Thallium and Other Elements (As,Cd, Cu,Mn, Pb,Se, Sb,and Zn) in Water and Sediment Samples from the Vicinity of the Zinc-Lead Smelter in Poland (3 pp)

Background, Goal and Scope   The aim of this study was to check the concentration of some elements in water samples collected near Pb-Zn mining and smelting works and comparison of the obtained data with results achieved for sediment samples originated from the same reservoirs. Objective   Here, 8 water samples and 3 bottom sediments collected from water reservoirs in the vicinity of 3 big Polish Pb-Zn smelters were analysed.Methods   Water analysis was performed after filtration through a 0.45 µm filter and pH adjustment to 2. For decomposition of dried sediment samples, microwave assisted digestion with total dissolution of silicate matrix was applied. The elements studied were determined using ICP-MS and ICP-OES methods. Results and Discussion   The concentrations of most studied elements in water samples were on the 0.X µg/L level, while only the contents of Tl, As, Mn, Cd, Pb and Sb in two water reservoirs were above the limits established for drinking water. The content of studied metals in sediments was in a wide range from X mg/kg (Se and Sb) to X000 mg/kg (Pb and Zn).Conclusions  and Recommendation. The study indicates that the impact of Tl, As, Cd, Cu, Mn, Pb, Zn, Se and Sb on bottom sediments is much more extensive than on the water in ponds located in the vicinity of a post-flotation waste heap. Monitoring of surface and underground waters, if limited only to the dissolved elements, can lead to faulty conclusions about environmental pollution. The bottom sediments mainly contain easily mobilised phases, and water-sediment equilibrium could be changed easily.     

Diversity of Ophiostomatales species associated with conifer-infesting beetles in the Western Carpathians

Bark beetles (Coleoptera: Curculionidae, Scolytinae) are commonly recognised as important agents of tree mortality in coniferous forests of the Western Carpathians. They, together with weevils, are consistently associated with ophiostomatoid fungi. Information regarding conifer beetle-associated fungi in the Western Carpathians remains incomplete and unreliable, particularly with respect to fir-infesting bark beetles. This study aims to clarify associations between fungi in the genera Graphilbum, Leptographium, Ophiostoma and Sporothrix (Ophiostomatales) and their beetle vectors in Norway spruce (Picea abies), European larch (Larix decidua) and silver fir (Abies alba). Samples associated with 20 bark beetle species and weevils were collected from nine stands in Poland and the Czech Republic. Fungi were isolated from adult beetles and galleries. Isolates were identified based on morphology, DNA sequence comparisons for four gene regions (ITS, LSU, ß-tubulin, TEF 1-α) and phylogenetic analyses. In total, 46 distinct taxa were identified, including 25 known and 21 currently unknown species. Several associations between fungi and subcortical insects were recorded for the first time. In addition, O. borealis and O. quercus were detected from A. alba for the first time. The composition of the fungal communities varied among the studied tree species and to a lesser degree among the beetle species. The spruce-infesting bark beetles were commonly associated with species of Leptographium s. l. and Ophiostoma s. str.; the larch-infesting bark beetles were often associated with Ophiostoma s. str. and Sporothrix, while the fir-infesting bark beetles were commonly associated with Ophiostoma s. str. and Graphilbum. The most commonly encountered fungal associates of the examined insects were (a) Grosmannia cucullata, G. piceiperda, Grosmannia sp. 1, Ophiostoma macroclavatum and O. piceae with the spruce-infesting bark beetles; (b) O. pseudocatenulatum and Sporothrix sp. 1 with the larch-infesting bark beetles; and (c) O. piceae, Ophiostoma sp. 2 and Graphilbum sp. 2 with the fir-infesting bark beetles. The differences in fungal associates among the bark beetle species occurring on P. abies, L. decidua and A. alba could be linked to the different habitats that these beetles occupy.     

Stocktake study of current fertilisation recommendations across Europe and discussion towards a more harmonised approach

The European Commission has set targets for a reduction in nutrient losses by at least 50% and a reduction in fertiliser use by at least 20% by 2030 while ensuring no deterioration in soil fertility. Within the mandate of the European Joint Programme EJP Soil ‘Towards climate-smart sustainable management of agricultural soils’, the objective of this study was to assess current fertilisation practices across Europe and discuss the potential for harmonisation of fertilisation methodologies as a strategy to reduce nutrient loss and overall fertiliser use. A stocktake study of current methods of delivering fertilisation advice took place across 23 European countries. The stocktake was in the form of a questionnaire, comprising 46 questions. Information was gathered on a large range of factors, including soil analysis methods, along with soil, crop and climatic factors taken into consideration within fertilisation calculations. The questionnaire was completed by experts, who are involved in compiling fertilisation recommendations within their country. Substantial differences exist in the content, format and delivery of fertilisation guidelines across Europe. The barriers, constraints and potential benefits of a harmonised approach to fertilisation across Europe are discussed. The general consensus from all participating countries was that harmonisation of fertilisation guidelines should be increased, but it was unclear in what format this could be achieved. Shared learning in the delivery and format of fertilisation guidelines and mechanisms to adhere to environmental legislation were viewed as being beneficial. However, it would be very difficult, if not impossible, to harmonise all soil test data and fertilisation methodologies at EU level due to diverse soil types and agro-ecosystem influences. Nevertheless, increased future collaboration, especially between neighbouring countries within the same environmental zone, was seen as potentially very beneficial. This study is unique in providing current detail on fertilisation practices across European countries in a side-by-side comparison. The gathered data can provide a baseline for the development of scientifically based EU policy targets for nutrient loss and soil fertility evaluation.     

Analysis of leaf surface sesquiterpenes in potato varieties

A comparative study of potato leaf sesquiterpenes was carried out. GC, GC-MS, and NMR analyses were used to identify and quantify the sesquiterpenes in the leaf surfaces of 10 potato (Solanum tuberosum) varieties. Two sesquiterpene alcohols and 17 sesquiterpene hydrocarbons were identified and quantitatively determined. The distribution of the sesquiterpenes was found to be variety-specific. The sesquiterpene contents of the different potato varieties were subjected to cluster and principal component analyses. The eight potato varieties of the main chemotype cluster were dominated by beta-caryophyllene (9-148 ng/cm2), germacrene D (2-46 ng/cm2), germacrene D-4-ol (0.4-31 ng/cm2), beta-sesquiphellandrene (1-34 ng/cm2), and an unknown sesquiterpene alcohol III (0.2-37 ng/cm2). Chemometric classification distinguished two varieties, Mila and Vistula, from a major cluster. The Vistula variety was distinguished from the others by its high contents of beta-caryophyllene, alpha-humulene, germacrene D, and germacrene D-4-ol and the Mila variety by beta-elemene, trans-alpha-bergamotene, (Z)-beta-farnesene, (E)-beta-farnesene, trans-beta-bergamotene, beta-sesquiphellandrene, and unknown sesquiterpene alcohols I, II, III.     

Availability of Microelements as Affected by Soil Reaction and Fertilization in Long-Term Field Experiments

Results presented in this paper come from two long-term fertilization experiments carried out in Skierniewice (since 1923) and _ yczyn (since 1960). The top layer of soil in Skierniewice contains a little more clay and silt (17%) than at _ yczyn (14%), but the climatic conditions are alike - the mean for many years shows the precipitation of 520mm and temperature 7,9°C. Soil samples for the determination of microelements were collected in the years 1999 and 2000 from the control plots (Ca) and from combinations corresponding to 3 fertilization systems: mineral (CaNPK), organic (Ca + FYM) and mixed organic-mineral (CaNPK + FYM). The contents of B, Cu, Zn, Mn were determined in 4 extract solutions: 0,01mol CaCl2, EDTA, 1mol HCl and HF (total contents). The paper presents average results for the last two years. The following amounts of microelements (%) in relation to the total contents in particular extract solutions were obtained: CaCl 2 m Cu 0,4< Zn 2,2< Mn 3,5 < B 5,0; EDTA - B 2,4 < Mn 5,7 < Zn 9,0 < Cu 13,9; HCl - B 7,8 < Cu 37,3 < Zn 37,6< Mn 72,5. The soil contents of readily soluble forms of Mn were significantly higher in non-limed than in limed soil, while the amounts of B, Cu and Zn were higher in limed soils as some amounts of Cu and Zn were present in lime fertilizers. In the system of mineral fertilization, some amounts of boron and zinc were introduced to the soil together with fertilizers, however, they were insufficient in view of nutritional requirements. On the other hand, in the organic-mineral system the amounts of introduced boron were adequate, copper and zinc twice higher and manganese four times higher. The use of mineral fertilization alone for 26 years, leads to soil depletion of zinc and copper and the use of manure alone enriches soil in those components.     

Effect of albumin and casein supplementation on the common carp Cyprinus carpio L. sperm motility parameters measured by CASA

Adhesion of sperm to a glass surface can be manifested by the low percentage of motile spermatozoa and lower speed. This crucial factor could disprove the sperm quality prediction from computer-assisted sperm analysis (CASA) results. In our study, this was particularly observed when solution L was used. However, protein supplementation was effective in the reduction in sperm adhesion. We found that albumin increased the sperm motility at concentrations of 0.25 % in solution L and 0.5 % in solution P. Casein was effective in both solutions at 0.25 % concentration. However, in solution L, the motility was lower than that measured in solutions P and L with the supplementation of BSA at 0.25 and 0.5 %. BSA supplementation was especially beneficial in terms of higher speed values regardless of an activation solution. Casein used with solution P allows us to obtain similar results as in the samples activated with BSA supplementation. Casein supplementation to solution L did not increase CASA results to the level observed in the samples activated with BSA added to the solution L. Our data suggest that the selection of an activation solution is one of the most crucial steps in the experiments requiring CASA data. Depending on the composition, different protein supplements should be used to reduce adhesion of sperm to a glass surface. We recommend the use of BSA at the 0.5–1 % concentration because at this level we obtained the highest CASA parameters in both activation media.     

Pharmacokinetic profiles of the analgesic flupirtine in dogs after the administration of four pharmaceutical formulations

ObjectiveFlupirtine (FLU) is a non-opioid analgesic with no antipyretic or anti-inflammatory effects which is used in the treatment of pain in humans. There is a substantial body of evidence on the efficacy of FLU in humans but this is inadequate for the recommendation of its off-label use in veterinary clinical practice. The aim of this study was to evaluate the pharmacokinetic profiles of FLU after intravenous (IV), oral immediate release (POIR), oral prolonged release (POPR) and rectal (RC) administrations in healthy dogs.Study designFour-treatment, single-dose, four-phase, unpaired, cross-over design (4 × 4 Latin-square).AnimalsSix adult Labrador dogs.MethodsAnimals in groups 1, 2 and 4 received a single dose of 5 mg kg⁻¹ FLU administered by IV, POIR and RC routes. Group 3 received a single dose of 200 mg subject⁻¹ via the POPR route. The wash-out periods were 1 week. Blood samples (1 mL) were collected at assigned times for 48 hours and plasma FLU concentrations were analysed by a validated HPLC method.ResultsAdverse effects including salivation, tremors and vomiting were noted in the IV group and resolved spontaneously within 10 minutes. These effects did not occur in the other groups. The FLU plasma concentrations were detectable in all of the treatment groups for 36 hours following administration. The pharmacokinetic profiles after extravascular administrations showed similar trends. The bioavailability values after POIR, POPR and RC were 41.93%, 36.78% and 29.43%, respectively. There were no significant differences in pharmacokinetic profiles between the POIR and POPR formulations. A 5 mg kg⁻¹ POIR dose or a 200 mg subject⁻¹ POPR dose gave plasma concentrations similar to those reported in humans after clinical dosing.Conclusion and clinical relevanceThis study provides pharmacokinetic data that can be used to design further studies to investigate FLU in dogs.     

Particulate selenium in the baltic sea atmosphere

In order to determine the concentrations of atmospheric selenium and divide it into size classes, aerosols were collected on nitro-cellulose filters, using the PIXE impactors, at three coastal stations in Preila (Lithuania), Hoburg (Gotland, Sweden) and Kap Arkona (Rugia Island, Germany). The determination of selenium and other elements in samples was made using the TXRF (total-reflection X-ray fluorescence) analysis. Air mass trajectories and the results of selenium studies indicate that the highest concentrations of selenium occur with air masses advecting from western and southwestern Europe, while the smallest concentrations were observed for air masses from north and northeastern Europe. It was confirmed that submicron particles of sizes smaller than 2 μm dominate the transport of suspended selenium. It was revealed that selenium in the air above the Baltic Sea is mainly of anthropogenic origin and originates in the same sources as the trace metals such as Cu, Pb and Zn.     

In situ weathering of rocks or aeolian silt deposition: key parameters for verifying parent material and pedogenesis in the Opawskie Mountains—a case study from SW Poland

Purpose

The aims: (1) to investigate the role of the in situ weathering of bedrock in providing substrate for soil formation; (2) to evaluate the aeolian contribution to the mountainous soils in the vicinity of thick loess cover; and (3) to determine the influence of aeolian silt on further soil development.

Materials and methods

The sampled sites were arranged along the slope toposequence, where an aeolian/silt admixture possibly occurred. Each soil catena started at the top of a hill and ended at its foot. Such an arrangement of the soil profiles ensured the tracking of loess thickness variations and detection of the depth of the residuum-derived materials. One reference soil profile, consisting of aeolian silt deposits, was made. The following soil properties were determined: pH, organic carbon content, soil texture, exchangeable acidity, exchangeable ions and geochemistry. In addition, thin sections were prepared from rock samples to confirm the type of bedrock present.

Results and discussion

The soils in the studied area were classified as Cambisols, Luvisols and Stagnosols, characterised by silt loam texture and a high content of elements indicating an aeolian silt contribution—Hf (7.4 to 14.8 ppm) and Zr (274.4 to 549.0 ppm). These values differ strongly from the residues typical of weathered quartzite, greywacke or catalasite substrates, which generally have low concentrations of Hf and Zr (0.7 to 7.0 ppm and 26.0 to 263 ppm, respectively). Based on the morphological, textural and geochemical data of the studied soils, three layers were distinguished, which show different inputs of aeolian silt: (1) an aeolian silt mantle; (2) a mixed zone in which loess was incorporated into the local material; and (3) a basal zone, free of the influence of aeolian silt. Based on the obtained results, a hypothetical pathway for soil formation in mountainous areas, influenced by aeolian silt admixing, was proposed.

Conclusions

Our study demonstrates that the soils developed in the Opawskie Mountains are characterised by an aeolian silt influence. This differentiates them from weakly developed soils, which comprise materials formed during in situ weathering only. Materials originating from bedrock weathering did not play an independent role as the parent material for the studied pedons. Aeolian silt was admixed with already existing autochthonous substrates, or completely replaced them. This influence on the soil formation resulted in the occurrence of Luvisols, Stagnosols and Cambisols. Such soils cannot be formed from the weathering of quartzites and greywackes, which contribute to a less structure-forming medium.