Ixodid ticks parasitizing Iberian red deer (Cervus elaphus hispanicus) and European wild boar (Sus scrofa) from Spain: geographical and temporal distribution

Commercial hunting of Spanish wild ungulates has made them an important economic resource. Wild ungulates may have an important role in the maintenance of ixodid tick populations, and also as reservoirs of pathogens. We studied the ixodid ticks that parasitize Iberian red deer and European wild boar from Spain. Ixodid ticks (n=6,336) were collected from 431 Iberian red deer and 142 wild boar in different regions of Spain. We found 10 different ixodid tick species parasitizing Iberian red deer, mainly Hyalomma marginatum marginatum (63.7%), Rhipicephalus (Boophilus) annulatus (7.9%) and R. bursa (7.5%). R. (Boophilus) annulatus was only collected in the province of Cádiz (southern Spain). We found 8 ixodid tick species on the wild boar, mainly Hy. m. marginatum (68.7%), R. bursa (14.6%) and Dermacentor marginatus (9.3%). We found one adult Hy. marginatum rufipes and one adult Hy. anatolicum excavatum parasitizing wild boar from south-central Spain. Mean prevalence of ixodid ticks was 41.3+/-0.08% (n=475) and 31+/-0.09% (n=284) and intensity of parasitization was 13.9+/-0.2 (n=283) and 13.6+/-0.3 (n=130) ticks/animal for Iberian red deer and wild boar, respectively. Only 5 of the 13 ixodid tick species found were shared by Iberian red deer and wild boar. This finding could indicate a host preference when Iberian red deer and wild boar share common habitats. In both Iberian red deer and wild boar from south-central Spain the monthly relative frequencies of Hy. m. marginatum and R. bursa presented an inverse pattern. The highest Hy. m. marginatum relative frequencies coincided with the lowest R. bursa relative frequencies along the year. R. bursa and I. ricinus were present in areas from northern to southern Spain while Hyalomma sp. and D. marginatus were exclusively collected in the two southern thirds of Spain. Haemaphysalis sp. and D. reticulatus were collected in northern Spain. Hy. m. marginatum and R. bursa were present during the whole year in red deer and wild boar from south-central Spain, showing more than one life cycle per year. These results are important for understanding the role of wild ungulates in the maintenance of tick infestations and to improve tick control programmes.     

Assay conditions and validation of a new UV spectrophotometric method using microplates for the determination of polyphenol content

A new spectrophotometric assay for the quantification of polyphenolic content has been validated. It is based on Prussian Blue method and adapted to microplate spectrophotometry. Prussian Blue reaction is critically dependent on reaction time, but microplate spectrophotometry permits exact measures of many samples at the same time. This new method is precise, reproducible, repeatable and exact.     

Toxicity of cultured bullseye puffer fish Sphoeroides annulatus

The toxin content in various life cycle stages of tank-cultivated bullseye puffer (Sphoeroides annulatus) were analyzed by mouse bioassay and ESI-MS spectrometry analysis. The presence of toxin content was determined in extracts of sperm, eggs, embryo, larvae, post-larvae, juvenile, pre-adult, and adult fish, as well as in food items used during the cultivation of the species. Our findings show that only the muscle of juveniles, the viscera of pre-adults, and muscle, liver, and gonad of adult specimens were slightly toxic (<1 mouse unit). Thus, cultivated S. annulatus, as occurs with other cultivated puffer fish species, does not represent a food safety risk to consumers. This is the first report of toxin analysis covering the complete life stages of a puffer fish under controlled conditions.     

Crop and stress coefficients in rainfed and deficit irrigation vineyards using sap flow techniques

Projected climate changes and expansion of viticulture to drier regions justify the installation and management of deficit irrigation (DI) strategies. Contradictory results on the effect of DI on crops may be ascribed to the incorrect application of these techniques. The lack of discrimination between basal crop (K _cb) and stress coefficients (K _s) can be an obstacle to proper irrigation management. A sap flow (SF) technique associated with microlysimeters and eddy covariance (EC) methods was applied to five commercial vineyards, aiming to discriminate those coefficients, during the driest period of the vegetative cycle. A comparative analysis of the coefficients, in relation to measured vegetation parameters (for K _cb) and plant water status (for K _s) is presented. K _cb, ranging from about 0.35 to 0.75, was highly correlated with leaf area index at stand level. K _s, which decreased till 0.2 in the most stressed vineyard, was well correlated to plant water status (K _s function), represented by predawn leaf water potential. K _s functions for the different experiments exhibited falling slopes with decreasing water status, with variable trends depending on the rates of maximal crop transpiration (T _m). These experimental results show that specific parameters for K _s functions, necessary to estimate water use and irrigation depths, in order to control the stress levels in DI scheduling, are also dependent on T _m.     

Response of stevia to nitrogen fertilization and harvesting regime in northeastern Portugal

The adaptation of stevia to the growing conditions of NE Portugal is assessed, including the tolerance of this species to cold temperatures, and the potential to produce biomass when grown as an annual crop and when subjected to various nitrogen (N) rates and two harvesting regimes. Almost all the plants died during the winter of 2014 (minimum temperatures peaked at ?8.0°C), making it necessary to replant the crop the following spring. With the best cutting regime (double cut) and N rate (150 kg N ha^?1), 1514.4 and 2390.0 kg ha^?1 of dry leaves were produced, respectively, in 2014 and 2015. Leaf chlorophyll concentrations estimated by the SPAD (Soil and Plant Analysis Development)-502 chlorophyll meter and a NDVI (Normalized Difference Vegetation Index) carried out by the Field Scout CM 1000 spectroradiometer showed significant differences among N rates, proving to be good indicators of plant N nutritional status. Based on the leaf analysis, provisional sufficiency ranges for N are proposed, namely 25–35 g kg^?1 for mid-summer and 15–25 g kg^?1 for early autumn. The fluorescence of chlorophyll a and the transient fluorescence intensity performed by the OS-30p+ fluorometer failed to show any stress induced by no-N control treatments in comparison to N-treated plants.     

Sheep fed with banana leaf hay reduce ruminal protozoa population

A ciliate protozoa suppression can reduce methane production increasing the energy efficiency utilization by ruminants. The physicochemical characteristics of rumen fluid and the profile of the rumen protozoa populations were evaluated for sheep fed banana leaf hay in replacement of the Cynodon dactylon cv. vaqueiro hay. A total of 30 male sheep were raised in intensive system during 15 days of adaptation and 63 days of experimental period. The animals were distributed in a completely randomized design that included six replicates of five treatments with replacement levels (0, 25, 50, 75, and 100%) of the grass vaquero for the banana leaf hay. Samples of fluid were collected directly from the rumen with sterile catheters. Color, odor, viscosity, and the methylene blue reduction potential (MBRP) were evaluated and pH estimated using a digital potentiometer. After decimal dilutions, counts of genus protozoa were performed in Sedgewick Rafter chambers. The averages of pH, MBRP, color, odor, and viscosity were not influenced by the inclusion of the banana leaf hay. However, the total number of protozoa and Entodinium spp. population significantly decreased at 75 and 100% inclusions of banana leaf hay as roughage.     

Intraspecific variation in growth and allocation patterns in seedlings of Pinus pinaster Ait. submitted to contrasting watering regimes: can water availability explain regional variation?

? Interpopulation variation in key functional traits of Pinus pinaster Ait. is well recognized. However, the relative importance of drought tolerance to explain this regional variation in the species remains elusive.

? Here, we raise the question whether water availability constitutes a likely driver of regional variation in biomass allocation, growth and morphological traits of ten populations that cover the distribution range of P. pinaster. We carried out an experiment where seedlings of five families per population were submitted to two contrasting watering treatments.

? The effects of water availability and population were significant for relative diameter and height growth rate, biomass allocation and number of lateral stems and dwarf shoots. Total dry mass significantly differed between watering treatments but it did not among populations. Populations could be clustered into four main groups. Root mass fraction explained most of the variation and significantly correlated to altitude but not to aridity.

? The geographical pattern of genetic variation found in morphology and biomass allocation did not translate into population differences in drought tolerance or phenotypic plasticity to water availability, indicating that water availability is not a likely driver of the regional variation observed in the studied traits of P. pinaster at the seedling stage.

     

Response of climate-growth relationships and water use efficiency to thinning in a Pinus nigra afforestation

Thinning is the main forestry measure to increase tree growth by reducing stand tree density and competition for resources. A thinning experiment was established in 1993 on a 32-year-old Pinus nigra Arn. stand in central Spain. The response of growth, climate-growth relationships and intrinsic water use efficiency (WUEi) to a stand density reduction were compared between moderate thinned plots and a control plot by a combined analysis of basal area increments (BAI), and C and O stable isotope ratios (δ¹³C_c and δ¹⁸O_c). BAI in the control plot showed a decreasing trend that was avoided by thinning in the thinned plot. Thinning also partially buffered tree-ring response to climate and trees were less sensitive to precipitation although more sensitive to temperature. Δ¹³C_c in the thinned plot was not modified indicating that stomatal conductance (g) and photosynthetic capacity (A) did not change or change in the same direction. However, δ¹⁸O_c decreased in the control plot (unrelated to δ¹⁸O of precipitation) but not in the thinned plot, suggesting a relative increase of temperature and irradiance and/or a decrease of air humidity after reducing the density consistent with an increase in A, g and BAI. As WUEi did not increase in the thinned plot, faster growth in this plot was caused by higher abundance of resources per tree. The trend of WUEi in both plots indicated low-moderate CO₂-induced improvements. Thinning might be a useful adaptation measure against climate change in these plantations reducing their vulnerability to droughts. However, because WUEi was not affected, the positive growth response might be limited if droughts and warming continue and certain thresholds are exceeded.     

Genetic structuring among populations of the great egret,Ardea alba egretta,in major Brazilian wetlands

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