Assessing foraging strategies of herbivores in Mediterranean oak woodlands: a review of key issues and selected methodologies

Montados are agro-silvo-pastoral ecosystems, typical of the Southwest Iberian Peninsula, of high socio-economic and conservation importance, where grazing is a dominant activity. Montados are characterized by an open tree canopy of Quercus sp. and a diverse undercover of shrubs and grasslands that constitute the plant food resources for grazing animals. Plant food resources of Montados are highly variable, both spatially and seasonally, in quantity and quality. Reliable and easy to use methods to monitor grazing are necessary to allow proper understanding of foraging strategies of grazing animals and to set sustainable grazing management. We describe the main characteristics of the plant food resources available for grazing animals, striking its variability, and revise the potential of using N-alkanes and saliva proteome methods to assess foraging strategies in Montados. In a scenario of dynamic multiple choices, we discuss the use of n-alkane methodology for the simultaneous estimation of diet composition and voluntary intake and saliva proteome as a mean of increasing the knowledge on diet adjustments.     

Plasma myeloperoxidase level and polymorphonuclear leukocyte activation in horses suffering from large intestinal obstruction requiring surgery: preliminary results.

Myeloperoxidase (MPO) is a specific enzyme of neutrophil azurophilic granules with a strong oxidative activity. Thanks to a radioimmunoassay of equine myeloperoxidase, the authors have observed a significantly higher plasma level of MPO in horses operated for strangulation obstruction of the large intestine (n = 6) than in horses suffering from a non-strangulating displacement of the large intestine (n = 9). For the 2 groups, 3 phases were distinguished: reception (P1), intensive care (P2) and terminal phase (P3). The mean peak values of MPO for these phases were 121.6 ng/mL (P1), 168.6 ng/mL (P2), and 107.0 ng/mL (P3) for the non-strangulating group, and 242.6 ng/mL (P1); 426.0 ng/mL (P2), and 379.5 ng/mL (P3) for the strangulation group. The variations of the mean peak values of plasma MPO were significantly different between the 2 groups and between the different phases. A significant increase of the least square means of MPO was observed between P1 and P2. A significant decrease of the least square means of the number of circulating leukocytes was observed between P1 and P3. Polymorphonuclear neutrophil activation could play a major role in the pathogenesis of acute abdominal disease and endotoxic shock.     

Identification of a Pro-Angiogenic Potential and Cellular Uptake Mechanism of a LMW Highly Sulfated Fraction of Fucoidan from Ascophyllum nodosum

Herein we investigate the structure/function relationships of fucoidans from Ascophyllum nodosum to analyze their pro-angiogenic effect and cellular uptake in native and glycosaminoglycan-free (GAG-free) human endothelial cells (HUVECs). Fucoidans are marine sulfated polysaccharides, which act as glycosaminoglycans mimetics. We hypothesized that the size and sulfation rate of fucoidans influence their ability to induce pro-angiogenic processes independently of GAGs. We collected two fractions of fucoidans, Low and Medium Molecular Weight Fucoidan (LMWF and MMWF, respectively) by size exclusion chromatography and characterized their composition (sulfate, fucose and uronic acid) by colorimetric measurement and Raman and FT-IR spectroscopy. The high affinities of fractionated fucoidans to heparin binding proteins were confirmed by Surface Plasmon Resonance. We evidenced that LMWF has a higher pro-angiogenic (2D-angiogenesis on Matrigel) and pro-migratory (Boyden chamber) potential on HUVECs, compared to MMWF. Interestingly, in a GAG-free HUVECs model, LMWF kept a pro-angiogenic potential. Finally, to evaluate the association of LMWF-induced biological effects and its cellular uptake, we analyzed by confocal microscopy the GAGs involvement in the internalization of a fluorescent LMWF. The fluorescent LMWF was mainly internalized through HUVEC clathrin-dependent endocytosis in which GAGs were partially involved. In conclusion, a better characterization of the relationships between the fucoidan structure and its pro-angiogenic potential in GAG-free endothelial cells was required to identify an adapted fucoidan to enhance vascular repair in ischemia.     

Resveratrol inhibits the activity of equine neutrophil myeloperoxidase by a direct interaction with the enzyme

Resveratrol is a polyphenolic antioxidant present in beverage and food known for its multiple protective effects. We report the inhibitory effects of resveratrol on equine myeloperoxidase (MPO), a hemic peroxidase present in the granules of the neutrophils involved in the inflammatory response. Resveratrol inhibited the production of reactive oxygen species (ROS) by stimulated equine neutrophils by acting as a direct scavenger of the ROS released by the cells but did not modify the degranulation of the stimulated neutrophils as the amounts of released MPO were unchanged. Resveratrol strongly inhibited the chlorination, oxidation, and nitration activities of MPO in a dose-dependent manner. By an original technique of specific immunological extraction followed by enzymatic detection (SIEFED), we demonstrated that resveratrol inhibited the peroxidasic activity of the MPO measured by a direct interaction such as the fixation of resveratrol on the enzyme. The observation of a decrease of the accumulation of compound II suggested that resveratrol acts as an electron donor for MPO reduction.     

Estimating effective population size of large marine populations,is it feasible?

Sustainable exploitation of marine populations is a challenging task relying on information about their current and past abundance. Fisheries‐related data can be scarce and unreliable making them unsuitable for quantitative modelling. One fishery independent method that has attracted attention in this context consists in estimating the effective population size (N_e), a concept founded in population genetics. We reviewed recent empirical studies on N_e and carried out a simulation study to evaluate the feasibility of estimating N_e in large fish populations with the currently available methods. The detailed review of 26 studies found that published empirical N_e values were very similar despite differences in species and total population sizes (N). Genetic simulations for an age‐structured fish population were carried out for a range of population and samples sizes, and N_e was estimated using the Linkage Disequilibrium method. The results showed that already for medium‐sized populations (1 million individuals) and common sample sizes (50 individuals), negative estimates were likely to occur which for real applications is commonly interpreted as indicating very large (infinite) N_e. Moreover, on average, N_e estimates were negatively biased. The simulations further indicated that around 1% of the total number of individuals might have to be sampled to ensure sufficiently precise estimates of N_e. For large marine populations, this implies rather large samples (several thousands to millions of individuals). If however such large samples were to be collected, many more population parameters than only N_e could be estimated.     

Histological localization assays for diflubenzuron in Pieris brassicae larva (Lepidoptera), by secondary ion microanalysis

No histochemical reaction is available to localize diflubenzuron in tissues. The insecticide can be localized in Pieris tissues by its fluoride atoms, used as tracers in secondary ion microanalysis, which is very sensitive for such light elements. In diflubenzuron-treated insects, especially in females, fluoride intensities are much higher than in controls; images demonstrate a wide distribution of the insecticide in the insect tissues. Thus, besides target organs, i.e., epidermis, diflubenzuron is present in other tissues, such as fat bodies, which may be related to the general metabolic effects and to a detoxification of the insecticide.     

Influence of oxalic acid on the adsorption of cadmium at the goethite surface

The adsorption/desorption of oxalic acid and Cd on goethite has been characterized in function of pH in KNO₃ medium. Electrophoretic mobilities and pH titrations show that while anion desorbs, cation adsorbs, the maxima of adsorption being in significantly different pH ranges. Adsorption-desorption phenomena of oxalic acid in function of pH are identical in absence or presence of Cd. Contrarily, not only adsorption, but also desorption of Cd is enhanced in presence of oxalic acid. Surface sites are shown to never be saturated even in presence of excess of ligand and cation. So, competition between anions and cations for surface sites is minor. Ligand in solution does not compete with surface sites for the Cd, probably due to the low tendency of oxalic acid to form complexes with Cd. Behavior of Cd in presence of oxalate is discussed and results suggest a surface-binding of Cd via an oxalate-bridge between the surface and the metallic cation.     

Effect of condensed tannin ingestion in sheep and goat parotid saliva proteome

Saliva appears as a defence mechanism, against potential negative effects of tannins, in some species of animals which have to deal with these plant secondary metabolites in their regular diets. This study was carried out to investigate changes in parotid saliva protein profiles of sheep (Ovis aries) and goats (Capra hircus), induced by condensed tannin ingestion. Five Merino sheep and five Serpentina goats were maintained on a quebracho tannin enriched diet for 10 days. Saliva was collected through catheters inserted on parotid ducts and salivary proteins were separated by two-dimensional gel electrophoresis. Matrix-assisted Laser desorption ionization - time of flight (MALDI-TOF) and liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to identify the proteins whose expression levels changed after tannin consumption. Although no new proteins appeared, quebracho tannin consumption increased saliva total protein concentration and produced changes in the proteome of both species. While some proteins were similarly altered in both species parotid salivary protein profile, sheep and goats also presented species-specific differences in response to tannin consumption.     

Failure of Lipopolysaccharides to Directly Trigger the Chemiluminescence Response of Isolated Equine Polymorphonuclear Leukocytes

Divergent results have been reported on the effects of lipopolysaccharides (LPS) on the activation of equine polymorphonuclear leukocytes (PMN). We therefore attempted to determine whether LPS alone can stimulate equine PMN or whether plasma factors are necessary. PMN were isolated from citrated blood on a discontinuous density gradient of Percoll. The luminol (10-3 mol/L)-enhanced chemiluminescence (CL) of 1.25 x 106 cells was measured after addition of Escherichia coli LPS (0.001-10 µg/ml) alone or after incubation in autologous plasma (1 h, 37°C). After direct stimulation with LPS, there were random variations of CL in 16 horses that were not reproducible from one sample to the next for the same horse. LPS which had been incubated in plasma gave a dose-dependent stimulation of the CL of the PMN, which did not occur if the plasma had been heat inactivated (1 h, 56°C). These results indicated a role for plasma factors, which were unlikely to be cytokines, as there were no monocytes or lymphocytes in the plasma incubated with LPS, but might have been complement fragments or LPS ligands, such as LPS binding protein. Studies using specific antibodies against these factors are needed to clarify this question.     

Artificially applied plant volatile organic compounds modify the behavior of a pest with no adverse effect on its natural enemies in the field

The use of volatile organic compounds (VOCs) derived from plants to manipulate insect pest behavior can be applied in an integrated pest management strategy (IPM) using a combination of attractive and repulsive stimuli. The “push–pull” strategy was developed on this idea in order to disturb and modify the distribution and abundance of pests to protect crops and reduce the use of agrochemicals. This field experiment investigates, in a “push–pull” context using broccoli as a target crop and Chinese cabbage as a pull component, the stimulo-deterrent effect of five synthetic VOCs (dimethyl disulfide, linalool, geraniol, eucalyptol and citronellol) on the oviposition of the cabbage root fly Delia radicum. With the exception of linalool, all compounds tested had a significant effect in the field and eucalyptol showed the most promising results, reducing oviposition on broccoli by 45 %. Moreover, eucalyptol was the only VOC able to reduce the final infestation of D. radicum, i.e., the number of pupae. The other VOCs reduced oviposition by 20–30 %. No adverse effect of the treatments was found on major parasitoids (Trybliographa rapae and Aleochara bipustulata) and potential predators of D. radicum. This study highlights the potential of VOCs as deterrent stimuli against agricultural pests in the field.