Phenotypic characterisation of the cellular immune infiltrate in placentas of cattle following experimental inoculation with Neospora caninum in late gestation

Despite Neospora caninum being a major cause of bovine abortion worldwide, its pathogenesis is not completely understood. Neospora infection stimulates host cell-mediated immune responses, which may be responsible for the placental damage leading to abortion. The aim of the current study was to characterize the placental immune response following an experimental inoculation of pregnant cattle with N. caninum tachyzoites at day 210 of gestation. Cows were culled at 14, 28, 42 and 56 days post inoculation (dpi). Placentomes were examined by immunohistochemistry using antibodies against macrophages, T-cell subsets (CD4, CD8 and γδ), NK cells and B cells. Macrophages were detected mainly at 14 days post inoculation. Inflammation was generally mild and mainly characterized by CD3⁺, CD4⁺ and γδ T-cells; whereas CD8⁺ and NK cells were less numerous. The immune cell repertoire observed in this study was similar to those seen in pregnant cattle challenged with N. caninum at early gestation. However, cellular infiltrates were less severe than those seen during first trimester Neospora infections. This may explain the milder clinical outcome observed when animals are infected late in gestation.     

Evaluation of a Kit for Estimating Organic Matter Concentrations in Bottom Soils of Aquaculture Ponds

Abstract The Hach CEL/700 Organic Matter Laboratory provided estimates of organic carbon concentrations in pond soil samples comparable to those obtained by the commonly-used Walkley-Black method. Dry ashing at 350–400 C tended to overestimate soil organic matter concentrations, and this procedure requires too much equipment and laboratory expertise for use on most aquaculture farms. The Hach kit can provide practical aquaculturists a simple and reliable method for measuring organic matter in pond soils. Samples from brackishwater ponds should be treated with mercuric sulfate to prevent chloride interference and overestimation of organic matter.     

Genotypic characterization and steviol glycoside quantification in a population of Stevia rebaudiana Bertoni from Paraguay

Journal of Crop Science and Biotechnology - Stevia rebaudiana (Bertoni) Bertoni is a widely grown species in various regions of the world, mainly due to its sweetening properties attributed to...     

Time course of endocrine changes in the hypophysis-gonad axis induced by hypobaric hypoxia in male rats

Chronic hypobaric hypoxia (CHH) induces a decrease in sperm output and spermatogenesis in male rats. The mechanisms that underlie these changes in testicular function are unknown and could involve changes in the hypophysis-gonad axis. We have tested the hypothesis that changes take place in the endocrine status (FSH, follicle stimulating hormone; LH, luteinizing hormone; testosterone) of rats subjected to CHH. Male Wistar rats were maintained under normobaric or hypobaric conditions (428 torr, 4,600 m). On days 0, 5, 15 and 30 post-exposure, 12 rats were anesthetized, their body weights were measured and blood samples were collected. The testicles were fixed in 4% formaldehyde and processed for histological analysis. In this time course, the FSH levels rose by day 5 post-exposure. On subsequent days, the FSH levels decreased in rats subjected to CHH with a tendency to remain higher than the normoxic group. The LH plasma levels decreased in rats exposed to CHH. Consistent with the decrease in LH levels, the plasma testosterone level decreased significantly after 30 days of CHH exposure. Integrated analysis of hormonal changes in rats subjected to CHH and the body dehydration that occurs in HH allows us to conclude that the effects of CHH on spermatogenesis may be partially related to changes in the hypophysis-gonad hormonal axis.     

Activity,molecular mass and hydrolysis on baker's yeast protein of extracellular proteases from the putative probiotic bacteria Microbacterium sp. strain 8L and Exiguobacterium mexicanum strain 8N

The bacteria Microbacterium sp. 8L and Exiguobacterium mexicanum 8N are known to improve the culture of Artemia franciscana using baker's yeast as food. Using spectrophotometry, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE), substrate‐SDS‐PAGE and pH‐stat in vitro‐digestibility assays, the activity, molecular mass and hydrolysis on baker's yeast protein of proteases from extracellular polymeric substances (EPS) of the strains 8L and 8N along with the pathogenic strains Microbacterium sp. 8R and Vibrio parahaemolyticus 588 CECT (Vp) were studied. The EPSs of 8L and 8R showed one activity band, on which the serine inhibitor phenylmethylsulphonyl fluoride (PMSF) had no effect. The EPSs of 8N showed four bands; two were unaffected by PMSF, whereas one was affected, and the other was partially affected. The EPSs of Vp showed two bands, one partially inhibited by PMSF. No inhibitory effects from 1‐chloro‐3‐tosylamido‐7‐amino‐2‐heptanone (trypsin inhibitor) were observed in the protease bands of the studied bacteria. The EPSs of 8L and 8N showed a similar degree of hydrolysis (pH‐stat). The EPSs of 8L had the lowest Dice index of similarity of yeast protein profiles at 1 h of reaction. We conclude that the strain 8L could benefit A. franciscana by providing bacterial proteases for digestion of baker's yeast.     

Main biotic drivers of tree growth in a developing Juniperus thurifera stand in central Spain

Over the last few decades, abandonment of traditional management practices in Spain has led to widespread stand densification and has favoured the expansion of some forest species that previously exhibited more restricted ranges. Spanish juniper (Juniperus thurifera L.) woodlands are experiencing this phenomenon due to agricultural land abandonment and a decrease in the livestock pressure. Yet the main drivers underlying stand structure and dynamics at this novel scenario are poorly understood. In this study, we investigate the main biotic drivers of tree growth in a high-density stand of the dioecious J. thurifera at an early developmental stage (mean tree age of 32, ca. 50 years after land abandonment). Tree growth was measured by coring 299 individuals of different reproductive classes (male, female and non-reproductive). Neighbourhood models were used to assess the relative importance of tree size and neighbourhood competition on tree growth of each reproductive class in the study plot. We found that tree size had the strongest effect on tree growth, whereas the effect of intraspecific competition was negligible. We observed differences in growth patterns among reproductive classes along trunk diameter sizes. Thus, at smaller sizes, the three reproductive classes presented identical patterns of growth. However, at bigger sizes, females were the fastest growing individuals, followed by males and non-reproductive individuals. Overall, our results suggest that in young J. thurifera monospecific forests, where self-thinning processes may have not undergone yet, tree size and the reproductive class could play a relatively more important role than competition as drivers of tree growth. These findings constitute new information which contributes to understanding growth dynamics at early developmental stages in this dioecious species. Furthermore, our results provide guidelines for silvicultural managing, suggesting that at these young juniper stands thinning would likely not translate into enhanced growth on remnant trees.     

Force-clamp spectroscopy monitors the folding trajectory of a single protein

We used force-clamp atomic force microscopy to measure the end-to-end length of the small protein ubiquitin during its folding reaction at the single-molecule level. Ubiquitin was first unfolded and extended at a high force, then the stretching force was quenched and protein folding was observed. The folding trajectories were continuous and marked by several distinct stages. The time taken to fold was dependent on the contour length of the unfolded protein and the stretching force applied during folding. The folding collapse was marked by large fluctuations in the end-to-end length of the protein, but these fluctuations vanished upon the final folding contraction. These direct observations of the complete folding trajectory of a protein provide a benchmark to determine the physical basis of the folding reaction.     

Effects of a diet enriched with eicosapentaenoic,docosahexaenoic and glutamine on cytokines as immunological markers for systemic inflammation in bitches before and after ovariohysterectomy

The post-operative period can generate immunological stress and can be modulated through supplementation with the omega-3 series of polyunsaturated fatty acids. This study aimed to evaluate the effects of diets enriched with high doses of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids and glutamine on inflammatory mediators in dogs before and after ovariohysterectomy (OVH). Twelve female dogs were divided into two groups: group A was fed a commercial diet without the addition of EPA and DHA, and group B was fed an experimental diet enriched with EPA and DHA (0.2 g/100 kcal). Experimental diet intake initiated 21 days before surgery and continued until 30 days after OVH. Parameters measured were serum cytokines (TNF-α, IL-6 and IL-10), C-reactive protein (CRP), IGF-1, lymphoproliferation and body composition before and after surgery. Statistical analyses were performed with SAS software considering the effects of age and diet and their interactions, and means were compared by the Tukey test. There was no difference between groups in body weight (p = .682), lean mass (p = .101) and body fat (p = .103). There were no group differences in serum concentrations of TNF-α, IL-6, IL-10, IGF-1, CRP and the percentage of lymphocyte proliferation. However, a time effect for TNF-α was observed (p < .001), in which T_0P (10 days after the surgical procedure) presented lower values of this cytokine when compared to the other evaluation time points; and interaction effects between group and time were observed for serum concentrations of IL-6 (p < .001) and IL-10 (p = .002). OVH procedure was not considered invasive enough to increase inflammatory cytokines after 30 days of surgery, as well as the dosage of the EPA and DHA used before and after the surgery did not modulate the inflammatory markers.     

Some chemical composition and biological activity of northern Argentine propolis

Twenty-five samples of propolis were collected from seven different regions in northern Argentina; ethanolic extracts of propolis were prepared from all samples, and the respective samples were examined for UV absorption spectra, RPHPTLC, RPHPLC, antimicrobial activity, antiradical activity, and total phenolic content. It was found that 16 of the 25 samples showed a phenolic profile similar to that found in samples from southern Brazil and corresponding to poplar-based propolis and that the rest of the samples showed a different profile and higher antimicrobial and antiradical activities.     

Temporal and spatial variations in dominance,diversity and biotic indices along a limestone stream receiving a trout farm effluent

Benthic macroinvertebrate response to organic pollution generated by a trout farm, located in the upper Rio Tajuña (Central Spain), was evaluated by calculating Whittaker's and Camargo's dominance indices; Margalef s, Menhinick's, Camargo's, Shannon's and MacArthur's diversity indices; and several biotic indices, including the Trent Biotic Index (TBI), the Chandler Biotic Score (CBS), the Biological Monitoring Working Party (BMWP), the Average Score Per Taxon (ASPT) and the Overall Quality Rating (OQR). An upstream sampling site (S-1) and three downstream stations placed 10 (S-2), 150 (S-3) and 1000 (S-4) m below the fish farm outlet were selected along the study area. Site S-3 was situated just below a man-made waterfall (1.8 m in height). Winter, spring and summer macrobenthic surveys were conducted during 1986. Plecopterans, ephemeropterans, coleopterans, trichopterans, amphipods and planarians decreased in abundance or were basent at downstream sampling sites, whereas simuliids, leeches, chironomids and tubificid worms increased in abundance. All diversity and biotic indices had highest and lowest values at S-1 and S-2, respectively, and higher values at S-3 (below the waterfall) than at S-4. However, Shannon's and MacArthur's diversity indices and the TBI were less effective in quantifying differences among sampling sites, exhibiting the lowest positive coefficients of linear correlation with species richness and other diversity and biotic indices. Both dominance indices had highest values at S-2, but similar values at S-1, S-3 and S-4. Biotic indices (except the TBI) and Margalef s, Menhinick's and Camargo's diversity indices appear to be the most sensitive measures for assessing macroinvertebrate response to changes in environmental conditions downstream from the fish farm. In addition, temporal variation in species richness, Margalef s index and biotic indices was similar, exhibiting their highest coefficients of variation at S-4. It is concluded that small waterfalls may improve the streamns self-purification of organic pollution generated by trout farms.