The Acidic Probe LysoSensor™ is not Useful for Acrosome Evaluation of Cryopreserved Ram Spermatozoa

To try new acrosomal probes for assessing ram spermatozoa, we compared the LysoSensor^? probe, which labels acidic organelles, with the frequently used peanut agglutinin acrosomal probe (PNA‐PE; phycoerythrin as fluorescent moiety). The previous microscopic observations showed a lack of relationship of LysoSensor^? with acrosomal status. Semen was obtained from five rams and frozen in four pools. Each pool was analysed carrying out a triple staining propidium ioide/PNA‐PE/LysoSensor^? Green DND‐189 to test acrosome labelling, and a double staining SYBR‐14/PI, to assess sperm viability. Stained samples were analysed by flow cytometry. All measurements were replicated. Data were processed using agreement and repeatability tests. LysoSensor^? labelling did not agree with PNA (mean of differences: 30.8%; coefficient of agreement: 22.6%), confirming microscopic observations. Nevertheless, when LysoSensor^? was compared with SYBR‐14/PI, the agreement was high (mean of differences: ?0.05%; coefficient of agreement: 5.07%). Repeatability of both methods was high and similar. LysoSensor^? did not seem to specifically stain the acrosome, but it may accumulate in the cytoplasm and label viable spermatozoa. Therefore, LysoSensor^? might not be used as an acrosomal probe in ram spermatozoa, but it could be used in other kind of studies, taking advantage of its pH sensitivity.     

Relating growth and nutrition to site factors in young chestnut plantations established on agricultural and forest land in northern Spain

The growth and nutritional status of 97 hybrid chestnut plantations on former agricultural land and forest land were studied in relation to site conditions (climatic variables and soil properties). A single family of curves was obtained for classifying the early height growth of hybrid and sweet chestnut. Soil limitations and nutrient deficiencies were evident in the former forest land, whereas the agricultural soils were found to be generally suitable for growing chestnut. The stands with highest growth rates were characterized by high foliar concentrations of K, P and Ca. Successful growth of the plantations was always related to high summer precipitation, low elevation and absence of long periods of risk of frost. The results confirm the adaptability of chestnut to most sites where it has been planted for timber production.     

Evaluation of pea protein concentrate as partial replacement of fish meal in practical diets for juvenile tench (Tinca tinca L.)

With the aim to evaluate different replacement levels of fish meal (FM) by pea protein concentrate (PPC) on survival, growth performance and body composition of juvenile tench (Tinca tinca), a 90‐day experiment was conducted with 6‐month‐old juveniles. Four practical diets (50% crude protein) differing in the level of replacement of FM protein by PPC protein were tested: 0% (control), 25%, 35% or 45%, corresponding to 0, 207.5, 290.4 or 373.3 g PPC kg^?1 diet respectively. Survival rates ranged from 96.4% to 98.5%. The 25% and 35% replacement diets resulted in similar growth values (P > 0.05) to those obtained with the control diet (average of the three feeding treatments: 57.57 mm total length, 2.48 g weight and 1.87% day^?1 specific growth rate). The 45% replacement diet had the lowest growth (P < 0.05). Fish with externally visible deformities ranged from 0% to 1.5%. The relation among amino acid profiles of the diets, body composition, growth performance of juveniles and amino acid requirements of other fish species is discussed. An amount of 290.4 g PPC kg^?1 diet (35% replacement of FM protein) can be included in juvenile tench diets without impairing growth performance.     

Feedbacks between vegetation pattern and resource loss dramatically decrease ecosystem resilience and restoration potential in a simple dryland model

Conceptual frameworks of dryland degradation commonly include ecohydrological feedbacks between landscape spatial organization and resource loss, so that decreasing cover and size of vegetation patches result in higher water and soil losses, which lead to further vegetation loss. However, the impacts of these feedbacks on dryland dynamics in response to external stress have barely been tested. Using a spatially-explicit model, we represented feedbacks between vegetation pattern and landscape resource loss by establishing a negative dependence of plant establishment on the connectivity of runoff-source areas (e.g., bare soils). We assessed the impact of various feedback strengths on the response of dryland ecosystems to changing external conditions. In general, for a given external pressure, these connectivity-mediated feedbacks decrease vegetation cover at equilibrium, which indicates a decrease in ecosystem resistance. Along a gradient of gradual increase of environmental pressure (e.g., aridity), the connectivity-mediated feedbacks decrease the amount of pressure required to cause a critical shift to a degraded state (ecosystem resilience). If environmental conditions improve, these feedbacks increase the pressure release needed to achieve the ecosystem recovery (restoration potential). The impact of these feedbacks on dryland response to external stress is markedly non-linear, which relies on the non-linear negative relationship between bare-soil connectivity and vegetation cover. Modelling studies on dryland vegetation dynamics not accounting for the connectivity-mediated feedbacks studied here may overestimate the resistance, resilience and restoration potential of drylands in response to environmental and human pressures. Our results also suggest that changes in vegetation pattern and associated hydrological connectivity may be more informative early-warning indicators of dryland degradation than changes in vegetation cover.     

Regulated deficit irrigation in ‘Clementina de Nules’ citrus trees. II: Vegetative growth

Summary

An experiment on Regulated Deficit Irrigation (RDI) was performed during 1995 and 1996 in an orchard planted with drip-irrigated ‘Clementina de Nules’/Carrizo Citrange in Moncada (Valencia) Spain. Treatments consisted of a control, irrigated during the whole year at 125% ET_lys and RDI treatments where irrigation was reduced to 25% or to 50% of crop evapotranspiration measured by a weighing lysimeter (ET_lys) during one of the following periods: I) flowering and fruit set (spring); II) initial fruit enlargement phase (summer) and III) final fruit growth and maturation phases (end of summer-autumn). An additional treatment, denominated 50%-Year, was irrigated at 50% ET_lys during the whole year. The effects of RDI treatments in relation to tree water status (pre-dawn Ψ^pd and midday Ψ^md leaf water potential, as well as their integral with time) show a good relation between total shoot emergence in the different growth flushes and the stress intensity reached (Ψ^pd) (r² = 0.80). This correlation was mainly due to the number of floral shoots (r² = 0.86) and not to vegetative ones (r² = 0.22). Similar results were observed between the stress integral at pre-dawn in each period and the former sprouting variables. In all cases, correlation was better with pre-dawn leaf water potential or with pre-dawn stress integral than with those at midday. RDI during spring reduced shoot length of the first growth flush (A1) and increased fruitlet fall after restarting normal irrigation. It also produced “off-season” flowering in the second flush growth (A2) and increased shoot emergence of the third flush growth (A3) with about 10% of them being floral. Summer RDI treatments did not alter vegetative growth, and although they produced off-season flowering (A3) it was much smaller than that of autumn RDI treatments, which in addition reduced vegetative growth with respect to the control. These effects, together with those of yield and fruit quality presented elsewhere, show that summer is the more appropriate period to apply RDI in “Clementina de Nules” mandarin trees.     

When structure means conservation: Effect of aggregate structure in controlling microbial responses to rewetting events

Rewetting events after a drought produce a pulse of soil respiration (the “Birch Effect”) that leads to a loss of carbon from soil, especially in Mediterranean ecosystems. Two main hypotheses have developed to explain the Birch effect: the “metabolic explanation”, based on the rapid consumption of intracellular osmolytes previously accumulated to survive to dry conditions, and the “physical explanation”, based on the consumption of carbon made accessible by physical destruction of internal structures of the soil.Here, we compared the respiration response of intact and crushed 9–4 mm aggregates from a California grassland soil under two different rewetting schemes: (1) successive short dry/wet events and (2) increased drought periods followed by a single rewetting. In intact aggregates, both microbial biomass and respiration rates were relatively stable through both experimental treatments. In crushed aggregates, through multiple short dry/wet cycles, both respiration rate and microbial biomass increased, while as drought length increased, biomass was unaffected but the magnitude of the following rewetting pulse increased. A mechanism that explains both these results is that crushing aggregates exposes occluded particular material that must be degraded into an immediately bioavailable form for microbes to take it up and metabolize it. Nitrification was generally higher in intact than crushed aggregates, suggesting the importance of physical association between nitrifiers and resources in regulating overall soil nitrification.This work suggests that physical processes are most important in driving respiration pulses through multiple rewetting cycles and that the physical association of organisms, substrates, and mineral particles are critical in controlling the functioning of the “microbial landscape”.     

Maize (Zea mays L.) from the Saharan oasis: adaptation to temperate areas and agronomic performance

Saharan maize had been adapted to extreme conditions and could have developed resistance to different stresses. However, genebanks and breeding collections have poor representation from Saharan germplasm and, particularly, from Algeria. This is a preliminary approach to investigate the adaptation and agronomic performance of a representative sample of Saharan maize. We evaluated open-pollinated Saharan populations along with European and American cultivars during 2?years in humid and dry Spanish locations and in Algiers (Algeria). Saharan populations were able to grow in temperate environments, although results were not consistent over years and the genotype-by- environment interactions were very important. Some of the Algerian populations evaluated in 2010 showed promising yield and anthesis silking interval over environments, but none of the Algerian populations evaluated in 2009 were adequately adapted to Spanish conditions. These results suggest that there are wide ranges of variability within Saharan maize for adaptation to temperate conditions, and further evaluations of Saharan maize should identify potential base populations for breeding maize in either side of the Mediterranean Sea. However, this germplasm requires prebreeding for adaptation to temperate conditions in order to be adequate for breeding programs in temperate areas.     

Effect of litter separation on 24-hour rhythmicity of plasma prolactin,follicle-stimulating hormone and luteinizing hormone levels in lactating rabbit does

Background  

This work describes the effect of a 48-h litter separation on 24-h patterns of plasma prolactin, FSH and LH concentration in female lactating rabbits kept under a 16:8 light-dark photoperiod (lights on at 0800 h).     

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.     

Aluminum speciation in the bulk and rhizospheric soil solution of the species colonizing an abandoned copper mine in Galicia (NW Spain)

Purpose  

The present study was carried out to identify and quantify the aluminum species present in the bulk and rhizospheric soil solution of the spontaneous vegetation colonizing the dump (Calluna vulgaris, Erica cinerea) and slope (C. vulgaris, E. cinerea, Salix atrocinerea) of an abandoned Cu mine in Touro (Galicia, NW Spain).