Expression of connexin 43 protein in testes, epididymides and prostates of stallions

REASONS FOR PERFORMING STUDY: Connexin 43 (Cx43) is a ubiquitously distributed gap junction protein in testes and other reproductive tissues. Adjacent cells share ions and small metabolites through intercellular channels, which are present in gap junctions. Previously, Cx43 has not been reported in testes, epididymides and prostates either in healthy stallions or cryptorchid horses. OBJECTIVES: To demonstrate the expression pattern of Cx43 in the reproductive tissues of stallions and examine whether naturally occurring bilateral cryptorchidism has any influence on distribution and expression of Cx43. METHODS: The expression and the presence of Cx43 protein were detected by means of immunohistochemistry and Western blot analysis using a polyclonal rabbit anti-Cx43 antibody. RESULTS: In stallions, gap junctions appeared as structures localised to cell-cell contacts between adjacent cells. In testes, Cx43 expression was detected in the interstitial tissue and seminiferous tubules, between Leydig and Sertoli, as well as Sertoli and germ cells. In epididymides, Cx43 was localised between epithelial cells, whereas in prostates, between secretory cells of the glandular epithelium. In the cryptorchid, a clear reduction of Cx43 signal was observed in all reproductive tissues. CONCLUSIONS: Coupling of Leydig cells via gap junctions may suggest that steroidogenic function of the testis is under the influence of these intercellular channels. Within seminiferous tubules, the expression was found to be stage-specific, pointing to its role in coordinating spermatogenesis. Differential distribution of Cx43 protein in the reproductive tract of normal and cryptorchid stallions indicates that expression is clearly dependent on the physiological status of the horse. POTENTIAL RELEVANCE: Detection of Cx43 expression in equine testicular, epididymal, and prostatic cells is important for a better understanding of the role of intercellular membrane channels in direct cell communication within the reproductive tract of stallions.     

Anti-oxidative/oxidative status of rat liver after ovariectomy

The lack of protective action of oestrogens which appears during menopausal period may be the reason of serious metabolic disturbances including oxidative stress. The hypothesis was stated that ovariectomy may induce the variations of antioxidant/oxidant status which can be detected in rat liver. A total of 102 healthy Wistar female rats were included in the experiment and divided into control (CON; n = 6), sham-operated (SHO; n = 48) and ovariectomized (OVX; n = 48) groups. Animals from SHO (n = 6) and OVX (n = 6) groups were killed every week during 8 weeks of experiment to detect dynamic changes in examined parameters. Anti-oxidative enzyme activities [glutathione peroxidase (GSH-Px); superoxide dismutase (SOD)] as well as total antioxidant capacity (TAC) and the intensity of lipid peroxidation, measured by the concentration of N,N,diethyl-p-phenylene diamine (DEPPD) radical, were determined in liver homogenates by the use of spectrophotometric methods. Wave-like patterns of examined parameters within 8 weeks of experiment were detected. GSH-Px activity tended to be higher in OVX animals and was significantly lower at 8th week when compared with 1st week of experiment. SOD activity was higher in SHO animals and showed significant differences between 3rd, 4th, 7th and 1st week. TAC values were significantly higher in OVX when compared with SHO groups in 2nd, 4th, 5th week and significantly lower in 3rd, 6th and 7th week of the experiment. The concentration of DEPPD radicals tended to increase in OVX group. In conclusion, ovariectomy which leads to oestrogen insufficiency is reflected as well in variation of anti-oxidative/oxidative parameters in rat liver homogenates.     

The culturable leaf mycobiome of Viscum album subsp. austriacum

In Poland's pine forests, mistletoe (Viscum album subsp. austriacum) abundance has increased over the past 20 years. This mistletoe infestation has decreased the growth and vigour of Scots pine trees. In this study, we surveyed the culturable fungi from healthy and diseased V. album subsp. austriacum leaves from two stands in Poland. In total, 63 distinct species were identified, 99.8% of which belonged to the phylum Ascomycota. The community compositions of fungi in mistletoe leaves were similar in healthy and diseased leaves as Alternaria alternata, Aureobasidium pullulans, Botryosphaeria visci, Fusarium paeoniae and Microsphaeropsis olivacea were consistently found in leaves of all symptom types. The most frequently isolated fungus from asymptomatic leaves was M. olivacea, followed by A. alternata, A. pullulans and Hypoxylon rubiginosum. In comparison, the most frequently isolated fungi from leaves with rusty-brown necrotic spots were (in decreasing order) M. olivacea, B. visci, F. paeoniae and A. alternata, while the most frequently isolated fungi from leaves with black or dark brown spots were M. olivacea, A. alternata, A. pullulans, Epicoccum layuense and F. paeoniae. This study was the first comprehensive report showing that certain fungal species may be pathogens of V. album subsp. austriacum in Poland. This study was also the first report of F. paeoniae, F. juglandicola, Diaporthe vacuae and Heterotruncatella spartii from V. album, and the first report of D. vacuae and H. spartii in Poland.     

Early development and allometric growth in hatchery‐reared Eurasian perch,Perca fluviatilis L.

The morphological development and allometric growth patterns of Eurasian perch, Perca fluviatilis L., a highly valued commercial species, were studied under intensive rearing conditions from hatching up to 50 DPH (Days Post Hatch). Based on the external morphology, four different phases during early development of Eurasian perch were identified: pre‐flexion larva 0–20 DPH (5.70–10.16 mm TL); flexion larva 22–30 DPH (11.09–15.14 mm TL) and post‐flexion larva/juvenile 32–50 DPH (18.00–24.75 mm TL). The results indicate that growth period when final replacement of all temporary (larval) structures and most important changes in the shape of P. fluviatilis occurred (between 13.95 and 24.06 mm TL, during flexion and post‐flexion phase) can be considered as a transitional period between the larva and juvenile. All body segments, except trunk length and tail length showed fast growth (positive allometry) throughout the entire studied period or up to the respective inflexion point with a common tendency to isometry. In addition, the specific behaviours (e.g. pelagic way of life) of Eurasian perch larvae resulted in some characteristic allometric growth patterns in the posterior region, different from the majority of other teleosts. The results are discussed with respect to the ontogeny of the functional morphology under both ecological and aquaculture considerations.     

The application of tannic acid to the elimination of egg stickiness at varied moments of the egg swelling process in pikeperch,Sander lucioperca (L.)

The aim of this article was to analyse the process of pikeperch, Sander lucioperca, egg swelling and to apply tannic acid to eliminate egg stickiness at different moments of the swelling process on artificially obtained eggs. The first experiment involved observation of egg swelling process and the second determined the effect of temperature (12, 14 and 16°C) on the egg swelling rate. The third experiment involved elimination of egg stickiness in a tannin solution (0.75 g L^?1) where eggs were submerged in a solution for 0.5, 1, 2 and 5 min – 5, 10, 15, 20, 25 and 30 min following gamete activation. The results indicate that the pikeperch egg swelling process lasts 30 min. It was found that the temperature did not affect the process duration. The results of the third experiment showed that the effectiveness of tannic acid application in egg stickiness elimination increases with time. The best result was obtained in groups of eggs submerged for 1 and 2 min (86.5% and 80.5% of larvae were obtained respectively) 30 min following the gamete activation. The results presented in this study for the first time indicate the possibility of highly effective procedure of egg stickiness elimination with tannic acid in pikeperch aquaculture.     

The effect of various nitrogen fertilization and foliar nutrition regimes on the concentrations of nitrates,ammonium ions,dry matter and N-total in carrot (Daucus carota L.) roots

Two field experiments (Experiment I in 2003–2005 and Experiment II in 2004–2005) with carrot c.v. ‘Kazan F₁’ were conducted at Trzciana village (50°06′N, 21°85′E). The experiments were arranged in a split-plot design with four replications. Two sub-blocks were identified in both experiments: I, without foliar nutrition; II, receiving plant foliar nutrition. The plants were sprayed three times alternately with: 2% urea solution, 1% solution of multi-component ‘Supervit R’ fertilizer (produced by Intermag, Poland) and again with 2% urea solution. Combinations with diversified nitrogen fertilization were distinguished within both sub-blocks. Experiment I comprised of: (1) Control, (2) Ca(NO₃)₂ 70, (3) Ca(NO₃)₂ 70 + 70, (4) (NH₄)₂SO₄ 70 and (5) (NH₄)₂SO₄ 70 + 70. Experiment II included: (1) Control, (2) ENTEC-26 35 + 35, (3) ENTEC-26 70 + 70, (4) ENTEC 26 105 + 105, (5) NH₄NO₃ 35 + 35, (6) NH₄NO₃ 70 + 70, (7) NH₄NO₃ 105 + 105. Where 70 kg N ha⁻¹ was used before sowing, whereas 35 + 35, 70 + 70 and 105 + 105 kg N ha⁻¹ were applied before sowing and as top dressing. Solid nitrogen fertilizer was added to the soil (produced by): Ca(NO₃)₂, Yara International ASA (Hydro); (NH₄)₂SO₄, Zak?ady Azotowe in Tarnów, Poland; NH₄NO₃, Zak?ady Azotowe in Pu?awy, Poland; and ENTEC-26, COMPO GmbH & Co. KG, Germany. The research aimed at determining the effect of diversified nitrogen fertilization and foliar nutrition on NO₃⁻, NH₄⁺, N-total and dry matter (d.m.) concentrations in carrot, and N uptake by storage roots. In Experiment I, nitrogen fertilization did not affect NO₃⁻ concentration, whereas in Experiment II, the applied N treatment increased NO₃⁻ concentration in carrot in relation to the control, except for the storage roots of plants fertilized with ENTEC-26 35 + 35. Nitrogen fertilization applied in both experiments caused a significant increase in N-total concentration in carrot and N uptake by storage roots in comparison with the control plants. In both experiments, nitrogen fertilization had a different effect on the concentrations of NH₄⁺ and d.m. in carrot. What is more, foliar nutrition treatments in both experiments had a different effect on the concentrations on NO₃⁻, N-total, d.m. in carrot and N uptake by carrot storage roots.     

Reproduction of chub, Leuciscus cephalus L., under controlled conditions

Research into artificial reproduction of chub was studied during three subsequent reproductive seasons. Chub reproduction was conducted in two different variations. In the first experiment different hormonal agents were given to fish, such as CPE, hCG, or an analogue of GnRH with dopamine antagonist (ovopel). In the second one, the optimum moment of applying of artificial reproduction was sought and the fish were only obtained an analogue of GnRH in different periods. The results obtained in two first seasons indicated that ovopel is the most effective agent for the artificial reproduction of chub. Consequently, it was decided that the fish would obtain only this agent in the final year of research. In last year (experiment II), chub spawners were taken to the hatchery under controlled conditions every few weeks from February to the beginning of June. Periodically, the maturity of oocytes was checked. As soon as the oocytes had achieved stage 2–3 of maturity, fish were obtained ovopel. It was noted that there was no problem with collecting semen. Obtaining oocytes was much more difficult. Most frequently, they were not obtained at all or obtained in very small amounts. It was also observed that the administration of hormones caused a very quick maturation of gonads and their over-maturation.     

Studying the Effects of Two Various Methods of Composting on the Degradation Levels of Polycyclic Aromatic Hydrocarbons (PAHs) in Sewage Sludge

The research comprised of studying the effect composting sewage sludge with sawdust and vermicomposting with earthworm Eisenia fetida has on the degradation of 16 polycyclic aromatic hydrocarbons (PAHs). Raw rural sewage sludge prior composting was more contaminated with PAHs than urban sewage sludge, in both cases exceeding EU cutoff limits of 6 mg/kg established for land application. Dibenzo[a,h]anthracene (DBahAnt), acenaphtylene (Acy) and indeno[1,2,3-c,d]pyrene (IPyr) were predominant in rural sewage sludge, whilst the urban sewage sludge contained the highest concentrations of benzo[b]fluoranthene (BbFl), benzo[k]fluoranthene (BkFl) and indeno[1,2,3-c,d]pyrene (IPyr). Thirty days of composting with sawdust has caused a significant reduction of 16 PAHs on average from 26.07 to 4.01 mg/kg (84.6%). During vermicomposting, total PAH concentration decreased on average from 15.5 to 2.37 mg/kg (84.7%). Vermicomposting caused full degradation of hydrocarbons containing 2 and 6 rings and significant reduction of PAHs with 3 aromatic rings (94.4%) as well as with 5 aromatic rings (83.2%). The lowest rate of degradation (64.4%) was observed for hydrocarbons with 4 aromatic rings such as fluoranthene, benzo(a)anthracene, chrysene and pyrene. On the other hand, the highest level of degradation was determined for PAHs with 2 rings (100%), 3 rings (88%) and 6 aromatic rings in the molecule (86.9%) after composting with sawdust. Acenaphthene and pyrene were found to be the most resistant to biodegradation during both composting methods.     

Effect of Preharvest Sprays of Calcium on Cracking and ‘Schattenmorelle’ Sour Cherry Fruit Quality Harvested Mechanically

The aim of the study was to examine effect of preharvest sprays of calcium (Ca) in the form of Ca-chloride (CaCl₂), Ca-nitrate [Ca(NO₃)₂], or a mixture of Ca-formate, Ca-acetate, CaCl₂, and Ca(NO₃)₂ on cracking and quality of ‘Schattenmorelle’ sour cherry fruit harvested mechanically. The experiment was conducted in 2008–2009 at a commercial orchard in central Poland. Mature trees grew on a coarse-textured soil poor in organic matter, at a spacing of 4.0 × 1.5 m. The spray treatments of Ca were performed at 7-day intervals, starting 28 days before harvest, at the rates of 5.0–5.6 kg Ca ha^?1 per season. The trees sprayed with water were treated as the control. Fruit were harvested mechanically when peduncle-fruit detachment force dropped below 3 N. The results showed that preharvest Ca sprays caused no leaf damage. This measure did not affect yield, mean fruit weight, soluble solids concentration and titratable acidity of fruit, and weight loss of fruit during 24 h after harvest. Fruit sprayed with Ca had improved status of this nutrient, and were less liable to juicy leakage from the stem scar, rain-induced cracking, and preharvest decay caused by Glomerella cingulata. The above effects of Ca sprays did not depend on the tested material. It was concluded that preharvest sprays of Ca as CaCl₂ and/or Ca(NO₃)₂ should be recommended in ‘Schattenmorelle’ sour cherry orchards to reduce fruit losses resulting from rain-induced cracking, leakage of juice, and the incidence of cherry bitter rot.     

Synthesis and phytotoxicity of new ionic liquids incorporating chiral cations and/or chiral anions

The aim of this work was to synthesize chiral ionic liquids as chiral solvents for organic synthesis and to evaluate the phyto(eco)toxicity of the new products and starting N-alkylimidazoles and their potential environmental influence on soil and plants. Chiral ionic liquids containing anions such as Cl-, Br-, TsO-, PF6(-), NO3(-), CF3SO3(-), and (+)- and (-)-C6H5CH(OH)C(O)O- were synthesized using (-)-(1R)-6,6-dimethylbicyclo[3.1.1]hept-2-ene-2-ethyl [(-)-(1R)-nopyl] halides (X = Cl, Br) and tosylate in 62-100% yields. The chloride 7 and the nitrate 13 ionic liquids possessed a toxicity dependent on the applied concentration. The lowest concentration causing a distinct reduction in plant germination/growth was 100 mg/kg. Spring barley better tolerated the ionic liquids (200 mg/kg) than common radish (100 mg/kg). The nitrate liquid did not exhibit an inhibiting effect on the germination ability of seeds. The starting N-methylimidazole used in lower concentrations (1 and 10 mg/kg of soil dry weight) was not phytotoxic, in contrast to higher doses (>1000 mg/kg).