Production of Hyaluronic Acid by Streptococcus zooepidemicus on Protein Substrates Obtained from Scyliorhinus canicula Discards

This work investigates the production of hyaluronic acid (H) by Streptococcus equi subsp. zooepidemicus in complex media formulated with peptones obtained from Scyliorhinus canicula viscera by-products. Initially, in batch cultures, the greatest productions were achieved using commercial media (3.03 g/L) followed by peptones from alcalase hydrolyzed viscera (2.32 g/L) and peptones from non-hydrolyzed viscera (2.26 g/L). An increase of between 12% and 15% was found in subsequent fed-batch cultures performed on waste peptones. Such organic nitrogen sources were shown to be an excellent low-cost substrate for microbial H, saving more than 50% of the nutrient costs.     

Effect of salicylic acid treatment on tomato plant physiology and tolerance to potato virus X infection

Salicylic acid (SA) is an inducer of systemic acquired resistance (SAR) and could be a potential candidate in the control of plant virus diseases. In this study we assayed under controlled conditions the potential effect of three doses of exogenous SA treatment on tomato plants infected with Potato virus X (PVX) and measured their effects on: different physiological parameters (gas exchange, stable isotopes, chlorophyll content), the activation of secondary metabolism, viral accumulation and induction of the expression of pathogenesis-related proteins (PRs) such as ß-1, 3-glucanase (PR2) and chitinase (PR3). SA treatment increased the expression of PR2, the activity of phenylalanine ammonia lyase (PAL) and the concentration of antioxidant compounds at 7 days post-treatment. Earlier expression of PR3 compared to PR2 was observed. SA treatment delayed the detection of PVX by ELISA in uninoculated leaves of mechanically infected tomato plants. Although the effect of PVX infection on physiological parameters was weak, moderate SA treatments showed enhanced photosynthesis, particularly for infected plants. The results obtained confirm that SA promotes major changes in the induction of resistance in tomato plants and suggest that treatment with exogenous SA could be considered to reduce the infections caused by PVX.     

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.     

Site-related influences on cone-borne inoculum and asymptomatic persistence of Diplodia shoot blight fungi on or in mature red pines

The shoot blight and canker pathogens Diplodia pinea and D. scrobiculata sporulate abundantly on cones of many pine hosts. Variation in incidence and abundance of potential inoculum from cones and frequency of asymptomatic persistence on or in shoots was examined for mature red pines in sites differing in dominant presettlement vegetation and soil type in Bayfield and Douglas counties in northern Wisconsin. Collections were made in each county from 6 plantations, 3 each in areas historically vegetated with jack pine and soils mapped as sands and three in areas historically vegetated with red pine with soils mapped as loamy sands. At each site, 5 cones were collected from each of 5 red pines and 10 shoots were collected from up to 5 red pines. Conidia from cones were quantified with a water wash and filtration technique. Diplodia species were cultured from surface-disinfested asymptomatic shoots. A species-specific PCR assay was used to identify the Diplodia species from cones and shoots. Although cones and asymptomatic shoots from each county yielded D. pinea and D. scrobiculata, D. pinea was detected more frequently. More conidia were obtained from cones from Douglas Co., where there is a history of severe shoot blight damage, than cones from Bayfield Co. In Douglas Co., more conidia were obtained from cones from plantations in areas of more sandy soil and presettlement jack pine dominance than cones from plantations in areas of less sandy soil and presettlement red pine dominance. The numbers of conidia and frequencies of cultural detection of Diplodia species from asymptomatic shoots at a site were positively correlated. These results provide evidence for site-related influences on abundance of pathogen inoculum and asymptomatic persistence on or in red pine crowns that may contribute to differences in frequency and severity of damage from Diplodia shoot blight.     

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.     

Environmental effects of a marine fish farm of gilthead seabream (Sparus aurata) in the NW Mediterranean Sea on water column and sediment

This study examined the effects of organic enrichment on water column, sediments and macrofauna caused by a fish farm in the Mediterranean Sea. Samples were collected on four sampling campaigns over a one‐year cycle. Significant differences were found in the water column in dissolved oxygen, dissolved inorganic nitrogen, phosphate and total phosphorus concentrations between the fish farm and the control. The increase in the dissolved inorganic nitrogen and phosphate concentrations at the fish farm modified the stoichiometric ratios between nutrients, with silicate acting as limiting nutrient at the fish farm 11% more than at the control. Nevertheless, chlorophyll a concentration in the water column was higher at the control station, probably due to the fouling of the underwater fish farm structures. Significant differences were found in sediment concentrations of organic matter, total phosphorus and redox potential between the fish farm and the control. The Canonical Correlation Analysis indicated that organic matter, total phosphorus, redox potential and% of gravels accounted for 68.9% of the total variance in the species data. Changes were observed in macrofauna, with a decrease in number of species and up to a nine‐fold increase in abundance with respect to the control.     

Exploring range shifts of contrasting tree species across a bioclimatic transition zone

Bioclimatic transition zones are supposed to encompass sensitive areas to global change effects on forest ecosystems. In this study, we attempt to detect shifts in the ranges of contrasting Iberian tree species in the submediterranean transition zone in Navarra, northern Spain. These shifts are analysed in the context of a significant increase in temperature over recent decades along with moderate land use changes. Data from national forest inventories (1971 and 2010) are compared through universal kriging (UK) and block kriging models to assess the shifts in the ranges of Quercus subpyrenaica, Quercus ilex, Pinus sylvestris and Fagus sylvatica. UK results predicted an increase in the presence probability of the four target species for the whole Navarra region. However, in the submediterranean zone, the presence probability of Q. subpyrenaica, P. sylvestris and F. sylvatica shows a shrinking trend, whereas Q. ilex is expanding its range, supporting a previous hypothesis of a “mediterranization” of this bioclimatic transition region. These trends are concomitant with recent elevational shift patterns towards higher elevations in the case of Q. subpyrenaica, P. sylvestris and F. sylvatica in the transition zone. Moreover, the expected increase in species richness as a consequence of geographical shifts and vegetation recovery is identified. The moderate human influence detected in the study area confirms the major role of climate warming as driver of species range shifts over the period. The results of this study highlight the suitability of bioclimatic transition zones for monitoring the effects of global change on natural ecosystems, providing evidences of the complex mechanisms affecting the distribution of forests.     

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.