Immunohistochemical distribution of antigens in liver of infected and immunized pigs with Ascaris suum

In the present work, we carry out an immunopathological study of the swine ascariosis, under different conditions (control, infection and immunization). Twenty-one Iberian pigs were used and divided in seven groups. Groups 1 and 2 were the uninfected and challenged controls, respectively. Groups 3 and 4 were weakly infected with increasing doses of Ascaris suum eggs and treated with pyrantel (Group 4). Groups 5-7 were immunized with 14, 42 and 97 kDa proteins from the parasite, respectively. Groups 2-7 were challenged with 10,000 infective eggs 7 days before the sacrifice. The focal parasitic granulomata with eosinophils and lymphocytes were the main histopathological lesions in the liver of reinfected pigs, while more marked cellular infiltrate and abundant connective tissue were seen in the livers of immunized animals. There were important deposits of antigens in the livers of immunized and infected pigs. Antigens were mainly located in the connective tissue, with positive staining detection of the somatic larvae antigen, the body wall from the adult worms and the 14-, 42- and 97-kDa proteins. However, cholangiols, biliary ducts and macrophages presented an immunohistochemical positive stain against excretory-secretory and somatic antigens from the larvae and the body fluid antigen from the adult parasite. The detection of A. suum antigens in the liver of infected pigs improves the diagnosis of swine ascariosis. It may be possible to apply these procedures for diagnosis of human ascariosis in liver biopsies since A. suum from swine have been previously used as a substitute for the study of the human parasite Ascaris lumbricoides.     

Cattle Drive Salmonella Infection in the Wildlife–Livestock Interface

The genus Salmonella is found throughout the world and is a potential pathogen for most vertebrates. It is also the most common cause of food‐borne illness in humans, and wildlife is an emerging source of food‐borne disease in humans due to the consumption of game meat. Wild boar is one of the most abundant European game species and these wild swine are known to be carriers of zoonotic and food‐borne pathogens such as Salmonella. Isolation of the pathogen, serotyping and molecular biology are necessary for elucidating epidemiological connections in multi‐host populations. Although disease management at population level can be addressed using a number of different strategies, such management is difficult in free‐living wildlife populations due to the lack of experience with the wildlife–livestock interface. Herein, we provide the results of a 4‐year Salmonella survey in sympatric populations of wild boar and cattle in the Ports de Tortosa i Beseit National Game Reserve (NE Spain). We also evaluated the effects of two management strategies, cattle removal and increased wild boar harvesting (i.e. by hunting and trapping), on the prevalence of the Salmonella serovar community. The serovars Meleagridis and Anatum were found to be shared by cattle and wild boar, a finding that was confirmed by 100% DNA similarity patterns using pulse field gel electrophoresis. Cattle removal was more efficient than the culling of wild boar as a means of reducing the prevalence of shared serotypes, which underlines the role of cattle as a reservoir of Salmonella for wild boar. To our knowledge, this is the first attempt to manage Salmonella in the wild, and the results have implications for management.     

Chemical constituents and antioxidant activity of sweet cherry at different ripening stages

The development and ripening process of sweet cherry (Prunus avium L. cv. 4-70) on the tree was evaluated. For this purpose, 14 different stages were selected in accordance with homogeneous size and color. Some parameters related to fruit quality, such as color, texture, sugars, organic acids, total antioxidant activity, total phenolic compounds, anthocyanins, and ascorbic acid were analyzed. The results revealed that in sweet cherry, the changes in skin color, glucose and fructose accumulation, and softening process are initiated at early developmental stages, coinciding with the fast increase in fruit size. Also, the decrease in color parameter a was correlated with the greatest accumulation of total anthocyanins. Ascorbic acid, total antioxidant activity (TAA), and total phenolic compounds decreased during the early stages of sweet cherry development but exponentially increased from stage 8, which coincided with the anthocyanin accumulation and fruit darkening. TAA showed positive correlations (r(2) = 0.99) with both ascorbic acid and total phenolic compounds and also with the anthocyanin concentration from stage 8. Taking into account the reduced shelf life of sweet cherry and to ensure that these fruits reach consumers with the maximum organoleptic, nutritional, and functional properties, it is advisable to harvest sweet cherries at stage 12 of ripening.     

The role of yellow lupin (Lupinus luteus) in the decline affecting oak agroforestry ecosystems

Phytophthora cinnamomi is a soilborne pathogen causing root rot in Mediterranean Quercus species growing in ‘dehesa’ rangeland ecosystems. Recently, it has been reported causing wilting and death of Lupinus luteus (yellow lupin), a spontaneous plant in southern Spain rangelands, but also frequently sowed for livestock grazing. In soils artificially infested with P. cinnamomi chlamydospores and planted with different cultivars of yellow lupin, a significant increase in the density of propagules was detected in comparison with the initial levels of inoculum and with the infested but not planted soil (control). In oak‐rangelands in which yellow lupine was planted, isolation and counting of colonies of P. cinnamomi from soil samples have shown the ability of this plant to maintain or even increase the inoculum density and thus facilitate the infection of trees. Results suggested that cultivation of yellow lupin in oak‐rangeland ecosystems should be avoided whether oak trees are affected by root disease caused by P. cinnamomi or not. This leguminous plant can act as an inoculum reservoir or even enhance inoculum soil levels available for oak root infections, exacerbating the oak decline severity in the region.     

Scarce population genetic differentiation but substantial spatiotemporal phenotypic variation of water-use efficiency in <Emphasis Type="Italic">Pinus sylvestris</Emphasis> at its western distribution range

Water and carbon fluxes in forests are largely related to leaf gas exchange physiology varying across spatiotemporal scales and modulated by plant responses to environmental cues. We quantified the relevance of genetic and phenotypic variation of intrinsic water-use efficiency (WUE_i, ratio of net photosynthesis to stomatal conductance of water) in Pinus sylvestris L. growing in the Iberian Peninsula as inferred from tree-ring carbon isotopes. Inter-population genetic variation, evaluated in a provenance trial comprising Spanish and German populations, was low and relevant only at continental scale. In contrast, phenotypic variation, evaluated in natural stands (at spatial level) and by tree-ring chronologies (at temporal inter-annual level), was important and ten- and threefold larger than the population genetic variance, respectively. These results points to preponderance of plastic responses dominating variability in WUE_i for this species. Spatial phenotypic variation in WUE_i correlated negatively with soil depth (r?=???0.66; p?<?0.01), while temporal phenotypic variation was mainly driven by summer precipitation. At the spatial level, WUE_i could be scaled-up to ecosystem-level WUE derived from remote sensing data by accounting for soil water-holding capacity (r?=?0.63; p?<?0.01). This outcome demonstrates a direct influence of the variation of leaf-level WUE_i on ecosystem water and carbon balance differentiation. Our findings highlight the contrasting importance of genetic variation (negligible) and plastic responses in WUE_i (large, with changes of up to 33% among sites) on determining carbon and water budgets at stand and ecosystem scales in a widespread conifer such as Pinus sylvestris.     

Susceptibility to Phytophthora cinnamomi of the commonest morphotypes of Holm oak in southern Spain

The four main morphotypes of Holm oak (Quercus ilex subsp. ballota) present in Andalusia (expansa, macrocarpa, microcarpa and rotundifolia) were infected with Phytophthora cinnamomi to determine their susceptibility to the root pathogen. No large differences were found among the four morphotypes in the infection of roots, which always showed a high degree of necrosis. However, the different responses of the foliage to infection separated the four morphotypes of Holm oak into three groups: very susceptible (microcarpa), susceptible (expansa) and moderately susceptible (rotundifolia and macrocarpa). The natural hybrid Q. ilex ballota–Q. faginea exhibited a low level of root and foliar symptoms when infected with P. cinnamomi. Quercus faginea could be considered as a source of resistance to P. cinnamomi in future breeding programmes.     

Management of a caseous lymphadenitis outbreak in a new Iberian ibex (Capra pyrenaica) stock reservoir

Background

In 2010, an Iberian ibex (Capra pyrenaica hispanica) stock reservoir was established for conservation purposes in north-eastern Spain. Eighteen ibexes were captured in the wild and housed in a 17 hectare enclosure. Once in captivity, a caseous lymphadenitis (CLA) outbreak occurred and ibex handlings were carried out at six-month intervals between 2010 and 2013 to perform health examinations and sampling. Treatment with a bacterin-based autovaccine and penicillin G benzatine was added during the third and subsequent handlings, when infection by Corynebacterium pseudotuberculosis was confirmed. Changes in lesion score, serum anti-C. pseudotuberculosis antibodies and haematological parameters were analyzed to assess captivity effects, disease emergence and treatment efficacy. Serum acute phase proteins (APP) Haptoglobin (Hp), Amyloid A (SAA) and Acid Soluble Glycoprotein (ASG) concentrations were also determined to evaluate their usefulness as indicators of clinical status.Once in captivity, 12 out of 14 ibexes (85.7%) seroconverted, preceding the emergence of clinical signs; moreover, TP, WBC, eosinophil and platelet cell counts increased while monocyte and basophil cell counts decreased. After treatment, casualties and fistulas disappeared and both packed cell volume (PCV) and haemoglobin concentration significantly increased. Hp, SAA and ASG values were under the limit of detection or showed no significant differences.

Conclusions

A role for captivity in contagion rate is suggested by the increase in antibody levels against C. pseudotuberculosis and the emergence of clinical signs. Although boosted by captivity, this is the first report of an outbreak of caseous lymphadenitis displaying high morbidity and mortality in wild ungulates. Treatment consisting of both vaccination and antibiotic therapy seemed to prevent mortality and alleviate disease severity, but was not reflected in the humoural response. Haematology and APP were not useful indicators in our study, perhaps due to the sampling frequency. Presumably endemic and irrelevant in the wild, this common disease of domestic small ruminants is complicating conservation efforts for the Iberian ibex in north-eastern Spain.     

Secreted proteases from Actinobacillus pleuropneumoniae serotype 1 degrade porcine gelatin, hemoglobin and immunoglobulin A.

It was found that 48 hour cultures of Actinobacillus pleuropneumoniae secreted proteases into the medium. Electrophoresis in polyacrylamide gels (10%) copolymerized with porcine gelatin (0.1%), of the 70% (NH4)2SO4 precipitate from the culture supernatants, displayed protease activities of different molecular weights: > 200, 200, 90, 80, 70 and 50 kDa. They had activity over a broad range of pHs (4-8), with an optimal pH of 6-7. All were inhibited by 10 mM EDTA, and reactivated by 10 mM calcium. They were stable at -20 degrees C for more than a month. The proteases also degraded porcine IgA and porcine, human, and bovine hemoglobin, although they appeared to be less active against the hemoglobins. The IgA was totally cleaved in 48 h, using supernatants concentrated with polyvinyl pyrrolidone or the 70% (NH4)2SO4. Extracellular proteases could play a role in virulence.     

Influence of surfactant supplementation and maceration on the feeding value of rice straw in growing-finishing diets for Holstein steers

Two trials were conducted to evaluate the interaction of the maceration process and surfactant (Tween 80) supplementation on feeding value of rice straw. Treatments were steam-flaked, corn-based diets containing 14% forage (DM basis), which was 1) Sudangrass hay; 2) ground rice straw; 3) ground rice straw plus 0.22% Tween 80; 4) macerated rice straw; and 5) macerated rice straw plus 0.22% Tween 80. In the maceration process, rice straw was passed through 2 sequentially placed pairs of corrugated rolls set at zero tolerance under a ram pressure of 62,050 millibars, similar to a conventional grain roller mill, except that the opposing rolls operated at different speeds (12 and 14 rpm, respectively). Sudangrass hay and rice straw (native and macerated) were ground through a 2.6-cm screen before incorporation into complete mixed diets. In trial 1, 125 Holstein steers (292 +/- 1.7 kg of BW) were used in a 188-d evaluation of the treatment effects on growth performance and carcass characteristics. In trial 2, 5 Holstein steers (224 +/- 3.5 kg of BW) with cannulas in the rumen and proximal duodenum were used in a 5 x 5 Latin square design to evaluate the treatment effects on digestion. There were no interactions between maceration and surfactant on growth or carcass characteristics. Tween 80 did not influence the feeding value of rice straw. Compared with grinding alone, maceration of rice straw increased the carcass-adjusted ADG (6%, P < 0.10), G:F (6%, P < 0.05), and dietary NE (5%, P < 0.05); DMI was similar across treatments. Assuming NE(m) and NE(g) of Sudangrass hay are 1.18 and 0.62 Mcal/kg, the NE(m) and NE(g) were 0.61 and 0.13 Mcal/kg for ground rice straw and 1.21 and 0.65 Mcal/kg for macerated rice straw. There were no treatment interactions on characteristics of digestion. Tween 80 did not influence ruminal or total tract digestion of OM, starch, NDF, or N. Compared with grinding alone, maceration of rice straw increased ruminal digestion of OM (7.7%, P < 0.10) and NDF (30.8%, P < 0.05), and total tract digestion of OM (2.3%, P < 0.10), NDF (21.1%, P < 0.01), and N (3.7%, P < 0.05). Total tract digestion of OM, NDF, starch, and N for the Sudangrass diet corresponded closely with that of the macerated rice straw diets. Maceration increases the feeding value of rice straw to a level similar to that of good-quality (flag stage of maturity) Sudangrass hay, which is attributable to increased OM and NDF digestion. Effects of surfactant supplementation on growth performance and digestion are not appreciable.     

Cereal protein analysis via Dumas method: Standardization of a micro-method using the EuroVector Elemental Analyser

The Dumas method (DM) of protein analysis in foods was standardized using two cereal references in order to identify the main sources of error during analysis; specifically, the study sought to establish which of the apparent critical factors (ACFs) were really significant influencing factors (IF) for the application of the DM to protein analysis in cereals.