Influence of foetal calf serum supplementation during in vitro embryo culture in Iberian red deer

Nowadays, the use of foetal calf serum (FCS) during in vitro embryo culture is very controversial. Whilst some authors have encouraged its use, others reject it because of its harmful effects. Although in vitro embryo production in red deer is a promising assisted reproductive technique, it is still in its infancy and a great effort is needed to update the protocols used. The aim of this study was to assess whether FCS supplementation in red deer embryo culture medium is necessary to produce blastocyst and, if so, when is the best time to add it in terms of blastocyst production and quality. In vitro blastocysts were cultured with FCS added at 24, 48 or 96 hours post‐insemination (hpi). In addition, a treatment without FCS was used as control. Six hundred and ninety‐four cumulus–oocyte complexes were collected for in vitro fertilization. Cleavage rate was examined at 48 hpi, and blastocyst yield was recorded on days 6, 7 and 8. FCS had no influence on cleavage and blastocyst rate for any of the treatments studied. However, the number of cells was higher (p = .025) in those blastocysts cultured with FCS from 48 hpi compared with FCS‐free culture media (93.88 ± 7.76 vs. 54.11 ± 8.36). In conclusion, the addition of FCS to the embryo culture medium at 48 hpi improves the quality of red deer blastocyst, although it does not affect the percentage of embryos obtained.     

Relationship between genealogical and microsatellite information characterizing losses of genetic variability: Empirical evidence from the rare Xalda sheep breed

Preservation of rare genetic stocks requires continual monitoring of populations to avoid losses of genetic variability. Genetic variability can be described using genealogical and molecular parameters characterizing variation in allelic frequencies over time and providing interesting information on differentiation that occurred after the foundation of a conservation program. Here we analyze the pedigree of the rare Xalda sheep breed (1851 individuals) and the polymorphism of 14 microsatellites in 239 Xalda individuals. Individuals were assigned to a base population (BP) or 4 different cohorts (from C1 to C4) according to their pedigree information. Genetic parameters were computed at a genealogical and molecular level, namely inbreeding (F), observed (H_o) and expected (H_e) heterozygosity, individual coancestry coefficients (f and f_m), average relatedness (AR), mean molecular kinship (Mk), average number of allele per locus (A), effective number of ancestors (f_a), effective population size (N_e and N_e(m)) and founder genome equivalents (N_g and N_g(m)). In general, the computed parameters increased with pedigree depth from BP to C4, especially for the genealogical information and molecular coancestry-based parameters (f_m, Mk and N_g(m)). However, H_o and H_e showed the highest values for C1 and the molecular heterozygote deficiency within population (F_IS(m)) showed the lowest value for C1, thus indicating that loss of genetic variability occurs very soon after the implementation of conservation strategies. Although no genealogical or molecular parameters are sufficient by themselves for monitoring populations at the beginning of a conservation program, our data suggests that coancestry-based parameters may be better criteria than those of inbreeding or homozygosity because of the rapid and strong correlation established between f and f_(m). However, the obtaining of molecular information in well-established conservation programs could not be justified, at least in economic terms.     

Antioxidant activities and metal acquisition in mycorrhizal plants growing in a heavy-metal multicontaminated soil amended with treated lignocellulosic agrowaste

The plant growth, nutrient acquisition, metal translocation and antioxidant activities [ascorbate peroxidase (APX), glutatione reductase (GR), superoxide dismutase (SOD) and catalase (CAT)] were measured in plants growing in a heavy-metal (HM) multicontaminated soil inoculated with selected autochthonous microorganisms [arbuscular mycorrhizal (AM) fungus and/or plant growth promoting bacteria (PGPB)] and/or amended with an Aspergillus niger-treated agrowaste. The treated agrowaste on its own increased root growth by 296% and shoot growth by 504% compared with non-treated control plants. Both chemical and biological treatments, particularly when combined, enhanced plant shoot and root development. The stimulation effect on plant biomass was concomitant with increased AM colonization, P and K assimilation, and reduced metal translocation from soil to plant shoot. The treated residue, particularly through interactions with AM inoculation, produced the expected bioremediation effect, leading to enhanced plant development and successful rehabilitation of contaminated soil. The enhancement of CAT, APX and GR activities caused by AM inoculation suggests that AM colonization helped plants to limit oxidative damage to biomolecules in response to metal stress. The response of the plant's antioxidant activities to the amendment appears to be related to enhanced plant biomass production. The application of amendments and/or microbial inoculations to enhance plant growth and reduce metal translocation in multicontaminated soil could be a promising strategy for remediating HM pollution.     

Contribution To Climate Change Of Forest Fires In Spain: Emissions And Loss Of Sequestration

ABSTRACT

Forest fires contribute to climate change mainly due to emission of greenhouse gases by biomass burning and loss of sequestration by sink destruction. The average contribution in Spain between 1998 and 2015 was 9,494,910 Mg CO_{2 eq} per year, 23.8% from biomass burning and 76.2% from loss of carbon sequestration, the latter three times higher than the former, although the emissions from combustion are usually the only accounted. Regarding to the vegetation burned, 43.6% of emissions come from forest (17.7% conifers, 4.8% hardwoods and 21.1% Eucalyptus), 53.7% from scrublands and 2.7% from grasslands. The loss of sequestration is 6.6% in the fire year and by 93.4% in previous years. Scrubland burning produces a greater amount of emissions than forests, but forest regeneration is slower, with greater influence on the loss of sequestration. It is essential a forest management focused on increase fire resilience and adaptation to climate change, increase the effectiveness of extinction works to reduce fire damages and implement actions to recover the burnt vegetation, because the loss of sinks is a critical aspect.     

Effect of Several Antioxidants on Thawed Ram Spermatozoa Submitted to 37°C up to Four Hours

Thawed ram spermatozoa were incubated at 37°C in the presence of dehydroascorbic acid (DHA), TEMPOL (TPL), N‐acetyl‐cysteine (NAC) and rutin (RUT), at 0.1 and 1 mm , in order to test their effects on sperm physiology. Cryopreserved spermatozoa from four rams were thawed, pooled, washed and incubated in TALP‐Hepes with 1 mm or 0.1 mm of each antioxidant, performing a replicate with induced oxidative stress (Fe²⁺/ascorbate). Motility (CASA), viability and mitochondrial membrane potential (flow cytometry) were analysed at 2 and 4 h. Lipoperoxidation (MDA production), intracellular reactive oxygen species (ROS) and DNA status (TUNEL) were analysed at 4 h. Antioxidants, except DHA 0.1 mm , decreased motility and kinematic parameters, but had little effect on viability or mitochondrial activity. Except 1 mm DHA, the antioxidants reduced ROS at 4 h. Moreover, NAC 1 mm , rutin and TEMPOL reduced ROS and DNA damage in the presence of oxidative stress. N‐acetyl‐cysteine, rutin 1 mm and TEMPOL reduced lipoperoxidation in the presence of oxidative stress. However, DHA did not affect lipoperoxidation. At 1 mm , DHA increased DNA damage in the absence of oxidative stress. Dehydroascorbic acid effects could arise from spermatozoa having a low capacity for reducing it to ascorbic acid, and it may be tested in the presence of other antioxidants or reducing power. Future research should focus in testing whether the inhibition of motility observed for NAC, rutin and TEMPOL is reversible. These antioxidants might be useful at lower temperatures (refrigerated storage or cryopreservation) when their protective effects could be advantageous.