Point mutations in BHV-1 Us3 gene abolish its ability to induce cytoskeletal changes in various cell types

The Us3 gene is conserved among alphaherpesviruses and codes for a protein kinase, a multifunctional protein involved in many phases of virus infection, like nuclear egress, modulation of apoptosis and modification of the cellular cytoskeleton. Bovine herpesvirus (BHV-1), a member of the Alphaherpesvirinae, contains an open reading frame homologous to Us3 of other herpesviruses, which has been identified as a serine/threonine kinase (Takashima, Y., Tamura, H., Xuan, X., Otsuka, H., 1999. Identification of the Us3 gene product of BHV-1 as a protein kinase and characterization of BHV-1 mutants of the Us3 gene. Virus Res. 59, 23–34). To study the activity of BHV-1 Us3, we have cloned its sequence under control of the human cytomegalovirus (HCMV) promoter/enhancer and introduced it into a recombinant baculovirus (Bac Us3). Confocal microscopy analysis showed profound cytoskeletal modifications in various BHV-1-permissive and non-permissive cells transduced with BacUs3. We observed that Us3 expression changed cellular shape and induced formation of long microtubule-containing cell projections, a phenomenon which had also been observed in cells expressing pseudorabies virus Us3. The intracellular localization of Us3 was mostly nuclear but when the protein accumulated it could be detected in the cytoplasm, cell membranes and projections. Mutated forms of BHV-1 Us3 with point mutations near or within the kinase catalytic domain did not affect cell morphology indicating that kinase activity of BHV-1 Us3 is required for its cytoskeleton remodelling function.     

Impact of diets with different proportions of linseed and sunflower oils on the growth, liver histology, immunological and chemical blood parameters, and proximate composition of pikeperch Sander lucioperca (L.)

The aim of the study was to determine the impact of applying different proportions of linseed (LO) and sunflower (SFO) oils in pikeperch diets on growth, histological changes in the liver, immunological and blood chemical parameters. The fish were fed isoenergetic and isoprotein feeds containing SFO (group 100SFO) or LO (group 100LO) in quantities of 67 g kg/feed, and a mixture of oils: 47 g SFO and 20 g LO kg/feed (group 70SFO/30LO) and 20 g SFO and 47 g LO kg/feed (group 30SFO/70LO). Dietary ratios of polyunsaturated fatty acids from the n-3 and n-6 series (n3/n6 index) were 0.36–2.15. Pikeperch were reared for 56 days in three replicates for each dietary treatment. Various dietary oils and ratios of n3/n6 did not impact fish growth, feed conversion ratio, viscerosomatic and hepatosomatic index, and size of the hepatocytes. Feeding the fish high quantities of LO and SO oils (groups 100LO and 100SFO) reduced the immunological response of the phagocytes and lymphocytes in the fish. Moreover, this resulted in significant differences among groups in the quantity of linolenic and linoleic acid in whole fish bodies, viscera, fillets, and livers. Various quantities of vegetable oils in the fish diets did not impact the quantity of arachidonic, eicosapentaenoic and docosahexaenoic acid in the fillets and livers. The immunological index and low quantities of linoleic acid in the fillets obtained in group 30SFO/70LO indicate that the n3/n6 dietary ratio of 1.35 was the most advantageous for feeding juvenile pikeperch feeds with vegetable oils.     

Development of the fungus Ceratocystis fimbriata in shoots of poplar clones with differing resistance1

Differences in the development and rate of spread of C. fimbriata hyphae between susceptible, moderately resistant and resistant clones were observed. Conidia of various shapes and ascospores appeared in necrotic parts of shoots of susceptible and moderately resistant clones while in the resistant one only cylindrical conidia were present.     

The effects of surgically implanted dummy tags on the survival,growth performance,and physiology of pikeperch (<Emphasis Type="Italic">Sander lucioperca</Emphasis>)

The aim of this work was to determine the impact of surgically implanted telemetry transmitters (TTs) on the growth, survival, hematological and biochemical indexes, and wound healing in juvenile pikeperch (Sander lucioperca) (body weight 60–90 g). Two incision suturing methods were used—silk sutures (experiment I—group ST) or tissue adhesive (experiment II—group GT). After tagging, the fish were held in a recirculating system for 35 days. No statistically significant differences were noted in the growth or condition indexes analyzed among the fish tagged with TT compared with those from the control groups (untagged). Substantial individual variability was noted, however, in the parameters examined in both the control and tagged groups. Among the hematological indexes, statistically significant differences were only noted in experiment I. Lower values of mean corpuscular volume and mean corpuscular hemoglobin were noted in group ST. Among the biochemical parameters, creatinine was statistically significantly threefold lower, magnesium and alkaline phosphatase (ALP) levels were lower, and ammonia levels were higher in group ST than in the control group. In experiment II, significant differences were only noted for ALP. Tissue adhesive was the superior and more effective method for closing the incision after TT implantation in juvenile pikeperch. This type of suturing facilitated faster healing, and it had less of an impact on juvenile pikeperch welfare.     

Meiotic gynogenesis affects the individual and gonadal development in sterlet Acipenser ruthenus

Larval growth and gonadal differentiation of gynogenetic sterlet Acipenser ruthenus were studied. The gynogenetic fish were produced by activation of albino sterlet eggs with UV‐irradiated sperm (collected from healthy coloration specimens), following by a heat shock of 34°C. Embryos were reared at 18.0 ± 0.5°C in a recirculation system and initially fed with Artemia sp. for 4 weeks and then fed with formulated feed. During the experiment, the length and weight of fish were recorded for the gynogenetic and control groups. Additionally, the analysis of gonadal sex differentiation was conducted with a 2‐week interval at 289, 303, 317, 331, 345 and 373 DPH. The mean body weight of the control group was significantly higher (p < .05) than the gynogenetic group only at 345 and 373 DPH, while the mean total body length was significantly higher at 331 and 345 DPH (p < .05). The gonad investigation was started at 289 DPH, and ~80% of females were found in the gynogenetic group during the trial period while the number of females and males was similar at 303, 331 and 373 DPH in the control group. The first symptoms of morphological differentiation appeared at 289 DPH, and the final shaping macroscopic structure of the gonad took place earlier in females than in males.     

Gamma-irradiation influence on the structure and properties of calcium caseinate-whey protein isolate based films. Part 2. Influence of polysaccharide addition and radiation treatment on the structure and functional properties of the films

The influence of gamma-irradiation (32 kGy) followed by the addition of polysaccharides (potato starch, soluble potato starch, and sodium alginate) and heating on the properties of the films based on calcium caseinate (CC)-whey proteins isolate (WPI) and the gels formed with CaCl(2) was evaluated. Radiation induced an improvement of the mechanical and barrier properties of all films. The polysaccharides' effect on the irradiated and non-irradiated CC-WPI gels could be predicted as the sum of their separate effects on CC and on WPI, apart from the alginate interaction with the irradiated CC-WPI. The better properties of the films achieved after admixing polysaccharides to the formerly irradiated protein solution correspond to the smaller strength of gels. Properties of the films and gels prepared using the irradiated proteins and alginate differed depending on whether alginate was admixed before or after irradiation. Results were related to the protein structure, interaction with polysaccharides, and the film's microstructure.     

Effect of Ectomycorrhiza on Cu and Pb Accumulation in Leaves and Roots of Silver Birch (Betula pendula Roth.) Seedlings Grown in Metal-Contaminated Soil

A pot experiment was conducted to study the effects of high concentrations of available Cu and Pb in soil originated from the vicinity of a copper foundry in Poland (Cu, 2,585–3,725 mg kg^?1 d.wt.; Pb, 1,459–1,812 mg kg^?1 d.wt.) on the growth and chemical constituents of Betula pendula seedlings. Control plants grew in unpolluted forest soil. Dry matter accumulation in the plants during the growing season and root/leaf mineral content were determined. Colonization of birch roots by ectomycorrhizal (ECM) fungi also was evaluated, as was soil dehydrogenase activity for influence of the metals on soil microorganisms. The heavy metals negatively affected seedling growth, ECM colonization, and soil dehydrogenase activity. A reverse relationship was found between ECM abundance and heavy metal concentrations in birch leaves, indicating the potential of mycorrhizas to protect the aboveground part of young silver birch seedlings from elevated environmental levels of Cu and Pb.     

Studies on the epiphytic bacteria on Populus tremula L. and Populus alba L. at the time of optimal and minimal cambium activity and infection by Venturia

The participation of epiphytic bacteria in the groups of bacteria dominating on the shoots and root collars of two year aspen and white poplar differs both in quantity and quality and depends on the chemical composition of the diffusion exudates related to cambium activity. This indicates that epiphytic bacteria can utilize carbon sources in the exudates and transform lignin precursors.     

Humic substances and aggregate stability in rhizospheric and non-rhizospheric soil

Purpose

The paper describes rhizospheric (Rs) and non-rhizospheric (nRs) soil to demonstrate the zone of the plant root impact on physical and chemical properties of the soil. The effects of the process accompanying the transformations of organic matter into humic substances in the rhizosphere of “common dandelion” Taraxacum officinale have been determined, and the properties of humic acids (HAs) were described. The importance of iron and clay minerals for the formation of a stable and water-resistant soil structure has been emphasized.

Materials and methods

The laboratory analysis involved determination of basic physical and chemical soil properties: texture, pH, cation exchange capacity (CEC), electrical conductivity, and content of total organic carbon (TOC) and dissolved organic carbon (DOC) and quality of humic substances: optical properties of HAs and its separation into hydrophilic (HIL) and hydrophobic (HOB) fractions, speciation of iron, glomalin operationally described as an easily extractable glomalin-related soil protein (EE-GRSP), and soil aggregate stability (SAS) of six size classes of soil aggregates.

Results and discussion

The Rs was reported with a higher TOC and DOC content (measured in the CaCl₂ extracts), however not significantly. The HAs isolated from Rs revealed a significantly higher content of humic substances at its initial decomposition stage, as compared with nRs. A significantly higher concentration of EE-GRSP was noted in the aggregates of the rhizospheric zone (mean 1.14 g kg^?1) than in the aggregates collected from root-free soil (mean 0.94 g kg^?1). There was noted the highest mean share of 1–3 mm soil aggregates in Rs as well as in nRs, respectively 44.4 and 38.3%. The soil material both in Rs and in nRs contained high amounts of exchangeable Ca²⁺, and smectite is the predominant clay mineral. It was favorable for the accumulation of organic carbon and for the formation of good soil physical condition (tilth). Higher but insignificant SAS values were observed for Rs (mean SAS?=?95.6%) than for nRs (mean SAS?=?93.9%).

Conclusions

The studies confirm the role of common dandelion roots in the process of organic carbon accumulation in rhizospheric zone and a favorable effect on the mechanism of the formation of water-resistant aggregates. Higher values of SAS for the Rs were affected by the content of TOC, DOC, exchangeable Ca²⁺ and the concentration of EE-GRSP, and, less considerably, the content of Fe and clay minerals.