Is the Production of Embryos in Small‐Scale Farming an Economically Feasible Enterprise?

The present assay attempts to evaluate the feasibility of using embryo transfer in small community farmers by in vivo study and by modelling the results obtained. From the total of 59 donor cows, 62.7% responded to treatment, with a significant difference (p = 0.002) in the percentage of the response between breeds, being 90.5% (19/21) in Holstein and 47.4% (18/38) in Brahman. A total of 283 embryos were graded as transferable, while 141 as non‐transferable, without difference in the percentage of transferable embryo by breed (p = 0.18). The mean of transferable embryos graded as class I and II was not different between Holstein and Brahman (p = 0.96 and p = 0.92, respectively); besides, no differences were observed in the other grades (non‐transferable). The highest difference in costs, regardless of its quality by breed, was seen in the lower levels of probable fertility of the embryo transferred, even reaching several hundred dollars. When modelling the expected costs for embryo produced and transferred, values can reach nearly $2000.00 when the probable fertility is only 10%. However, when the probable fertility was 60%, embryo cost was close to $300.00. This technology seems to be viable on average or high‐scale systems, having a superovulatory response between 60 and 80% with 4–6 transferrable embryos. Yet, in small‐scale farming, due to the reduced number of donors and/or recipients, the costs surpass the economical feasibility of the technique.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Protein Composition of Seminal Plasma in Fractionated Stallion Ejaculates

Seminal plasma (SP) contains several types of compounds derived from the epididymides and accessory glands. The aim of this study was to examine the protein composition of different ejaculate fractions. Trial I: fractionated ejaculates were collected from two normal and two subfertile stallions. Samples containing pre‐sperm fluid and the first sperm‐rich jets (HIGH‐1), the main sperm‐rich portion (HIGH‐2), the jets with low sperm concentrations (LOW), and a combined whole‐ejaculate (WE) sample was centrifuged, and the SP was filtered and frozen. A part of each SP sample was stored (5°C, 24 h) with spermatozoa from HIGH‐2 and skim milk extender. Sperm motility was evaluated after storage in extender mixed with the stallion’s own SP or SP from one of the other stallions (sperm from a normal stallion stored in SP from a subfertile stallion and vice versa). Protein composition was analysed using reverse‐phase liquid chromatography (RP‐HPLC), N‐terminal sequencing and mass spectrometry. The area‐under‐the‐curve (AUC) was used for quantitative comparison of proteins within fractions. Trial II: semen samples were collected from seven stallions. Fractions with the highest (HIGH) and lowest (LOW) sperm concentrations and WE samples were examined using SDS‐PAGE and densitometry. No significant differences emerged between fractions in the AUC‐values of the Horse Seminal Protein‐1 (HSP‐1) and HSP‐2 peaks, or the peak containing HSP‐3 and HSP‐4 (HSP‐3/4). Levels of HSP‐1, HSP‐2 and HSP‐3/4 were not significantly correlated with total sperm motility, progressive sperm motility or average path velocity after storage. Significant differences between ejaculate fractions in the amount of different protein groups present in SP were not found in Trial I; but in Trial II, the proteins in the 60–70 kDa range were more abundant in LOW than in HIGH and WE, indicating that this band contained proteins derived mainly from the seminal vesicles, which produce most of the SP in LOW.     

Fungi occurring on forests injured by air pollutants in the Upper Silesia and Cracow industrial regions. X. Mycoflora of dying young trees of Alnus incana

The results of a 10-year study on the mortality of Alnus incana in the Upper Silesia industrial region (Poland) are presented in this paper. Fungi which infected the trees at different stages of the disease were identified. The most common were: Cryptospora suffusa, Hypoxylon fuscum, Peniophora cinerea, P. erikssonii, Pezicula cinnamomea, Tympanis alnea, and Valsa diatrypa.     

Oak decline: I. Fungi associated with various disease symptoms on overground portions of middle-aged and old oak (Quercus robur L.)

Fungi colonizing overground portions of Quercus robur art presented separately for each of the following types of disease symptoms: leaves with atrophy and necrotic areas, dead branches, dead tops of branches, dead water sprouts, local necrotic areas in bark and phloem, locally dead and discoloured sapwood in debarked places, local discolorations inside sapwood, extensive nec-rotic areas on stems, dead trees. Over 80 species of fungi were found in all, mainly from As-comycotina and Deuteromycotina. Connected with each type of symptom there were several species of fungi which were more frequent than the remaining ones. In most cases fungi seem to be of secondary importance in development of symptoms characteristic for oak decline.     

Cryptic speciation in Hymenoscyphus albidus

Ash dieback caused by the mitosporic ascomycete Chalara fraxinea is a novel disease of major concern affecting Fraxinus excelsior and Fraxinus angustifolia in large parts of Europe. Recently, its teleomorph was detected and assigned to Hymenoscyphus albidus, which has been known from Europe since 1851. In this study, we present molecular evidence for the existence of two morphologically very similar taxa, H. albidus, which is lectotypified and Hymenoscyphus pseudoalbidus sp. nov. Differences were found between the species in the loci calmodulin, translation elongation factor 1‐α and the internal transcribed spacers of the rDNA genes, and strong differentiation was obtained with ISSR markers. It is likely that H. albidus is a non‐pathogenic species, whereas H. pseudoalbidus is a virulent species causing ash dieback. Genotyping herbarium specimens showed that H. pseudoalbidus has been present in Switzerland for at least 30 years prior to the outbreak of the epidemic.     

Post-fire soil respiration in relation to burnt wood management in a Mediterranean mountain ecosystem

After a wildfire, the management of burnt wood may determine microclimatic conditions and microbiological activity with the potential to affect soil respiration. To experimentally analyze the effect on soil respiration, we manipulated a recently burned pine forest in a Mediterranean mountain (Sierra Nevada National and Natural Park, SE Spain). Three representative treatments of post-fire burnt wood management were established at two elevations: (1) “salvage logging” (SL), where all trees were cut, trunks removed, and branches chipped; (2) “non-intervention” (NI), leaving all burnt trees standing; and (3) “cut plus lopping” (CL), a treatment where burnt trees were felled, with the main branches lopped off, but left in situ partially covering the ground surface. Seasonal measurements were carried out over the course of two years. In addition, we performed continuous diurnal campaigns and an irrigation experiment to ascertain the roles of soil temperature and moisture in determining CO₂ fluxes across treatments. Soil CO₂ fluxes were highest in CL (average of 3.34 ± 0.19 μmol m⁻² s⁻¹) and the lowest in SL (2.21 ± 0.11 μmol m⁻² s⁻¹). Across seasons, basal values were registered during summer (average of 1.46 ± 0.04 μmol m⁻² s⁻¹), but increased during the humid seasons (up to 10.07 ± 1.08 μmol m⁻² s⁻¹ in spring in CL). Seasonal and treatment patterns were consistent at the two elevations (1477 and 2317 m a.s.l.), although respiration was half as high at the higher altitude.Respiration was mainly controlled by soil moisture. Watering during the summer drought boosted CO₂ effluxes (up to 37 ± 6 μmol m⁻² s⁻¹ just after water addition), which then decreased to basal values as the soil dried. About 64% of CO₂ emissions during the first 24 h could be attributed to the degasification of soil pores, with the rest likely related to biological processes. The patterns of CO₂ effluxes under experimental watering were similar to the seasonal tendencies, with the highest pulse in CL. Temperature, however, had a weak effect on soil respiration, with Q₁₀ values of ca. 1 across seasons and soil moisture conditions. These results represent a first step towards illustrating the effects of post-fire burnt wood management on soil respiration, and eventually carbon sequestration.     

Optimisation of sodium and potassium concentrations and pH in the artificial seminal plasma of common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.

The effect of sodium and potassium concentrations as well as optimal pH on the motility of common carp Cyprinus carpio L. sperm during short-term storage in artificial seminal plasma (ASP) was investigated. Sperm was collected from individual males (n?=?5) and each sample diluted tenfold (1:9) in ASP (sperm:extender) containing 2 mM CaCl₂, 1 mM Mg₂SO₄ and 20 mM Tris at pH 8.0 and supplemented by the following concentrations of sodium and potassium (mM/mM): 0/150, 20/130, 40/110, 75/75, 110/40, 130/20 and 150/0. The osmolality of all ASP variants was set at 310 mOsm kg^?1. Sperm motility was measured using a CASA system during 72 h of storage. Immediately after dilution, sperm motility was high (90%) both in each variant and in the control group (fresh sperm). After 72-h storage, the highest sperm motility was noted in ASP containing 110 mM NaCl and 40 mM KCl. No differences were found in the motility of samples preserved within the pH range of 7.0–9.0. Our data suggest that for the short-term storage of common carp sperm, whereas the pH of the solution does not play a crucial role, a specific potassium concentration of around 40 mM is required.