Determination of arylsulphatase and phosphatase enzyme activities in soil using screen-printed electrodes modified with multi-walled carbon nanotubes

Sustainability of agricultural systems has become an important issue all over the world. The activity of enzymes is potentially an important quality bioindicator in soils. The aim of the present study was to develop a simple and convenient assay to determine the activity of arylsulphatase (AS), acid (ACP) and alkaline phosphatase (ALP) in agricultural soil. The activities of these enzymes were detected using a non-electroactive substrate, which produces an electroactive product. To this end, p-aminophenyl phosphate (pAPP) was used as a substrate which is converted to p-aminophenol (pAP) after enzymatic dephosphorylation; and 4-nitrocatechol sulphate (4-NCS) was used as a substrate for AS activity based on its catalytic effect on the hydrolysis of 4-NCS into 4-nitrocatechol (4-NC). The products of both enzymatic reactions were quantified on carbon-based screen-printed electrodes (SPCEs) modified with carbon nanotubes (CNTs), using Osteryoung square-wave voltammetry (OSWV). The determination of the reaction products allowed more sensitive determination of ALP, ACP and AS activities in soil than that obtained with a spectrophotometric method. This assay also diminishes the generation of waste, which is desirable in green analytical chemistry. The optimization of the analytical procedure in terms of the nature of electrode type, applied potential, pH of solution, and precision of measurements is reported.     

Differential contribution of two Ppd-1 homoeoalleles to early-flowering phenotype in Nepalese and Japanese varieties of common wheat

Wheat landraces carry abundant genetic variation in heading and flowering times. Here, we studied flowering-related traits of two Nepalese varieties, KU-4770 and KU-180 and a Japanese wheat cultivar, Shiroganekomugi (SGK). These three wheat varieties showed similar flowering time in a common garden experiment. In total, five significant quantitative trait loci (QTLs) for three examined traits, the heading, flowering and maturation times, were detected using an F₂ population of SGK/KU-4770. The QTLs were found at the Ppd-1 loci on chromosomes 2B and 2D and the 2B QTL was also confirmed in another F₂ population of SGK/KU-180. The Ppd-D1 allele from SGK and the Ppd-B1 alleles from the two Nepalese varieties might be causal for early-flowering phenotype. The SGK Ppd-D1 allele contained a 2-kb deletion in the 5′ upstream region, indicating a photoperiod-insensitive Ppd-D1a allele. Real-time PCR analysis estimating the Ppd-B1 copy number revealed that the two Nepalese varieties included two intact Ppd-B1 copies, putatively resulting in photoperiod insensitivity and an early-flowering phenotype. The two photoperiod-insensitive Ppd-1 homoeoalleles could independently contribute to segregation of early-flowering individuals in the two F₂ populations. Therefore, wheat landraces are genetic resources for discovery of alleles useful for improving wheat heading or flowering times.     

Effect of restricted suckling on milk yield,composition and flow,udder health,and postpartum anoestrus in grazing Holstein cows

The effect of restricted suckling on milk yield and composition, udder health, and postpartum anoestrus in dairy cows in pasture-based systems, was studied in 32 Holstein multiparous cows and their calves. At calving, each cow–calf pair was randomly assigned to one of two treatments: restricted suckling (RS) of the cows by her own or another calf, twice daily for 30 min or artificial rearing (AR) of the calves with milk obtained from the bulk tank, offered twice a day in buckets. Treatments were applied until week 8 after calving. The diet of the cows consisted of direct grazing in improved pastures, corn silage and a commercial concentrate which was offered at milking. Milk production and composition, udder health, body condition score of the cows, body weight and milk intake of the calves were measured weekly, and the first postpartum ovulation was determined three times a week by ovarian ultrasonography. Cows with RS management had a lower machine-milked milk yield (17.9 vs. 24.8 kg/d), a lower fat percentage (3.21 vs. 4.11%) and 4% fat-corrected milk yield (16.2 vs. 25.7 kg/d), and also a lower average milk flow (1.35 vs. 1.76 kg/min) than cows in the AR treatment. There was no effect of treatment on milk protein percentage or udder health as measured by milk electrical conductivity. The interval from calving to first postpartum ovulation was shorter in the AR cows than in the RS cows (18.5 vs. 21.8 days). The RS calves consumed more milk (7.2 vs. 5.4 kg/d), gained more body weight (0.813 vs. 0.656 kg/d), and had a higher body weight at weaning (84.3 vs. 73.3 kg) than AR calves. Restricted suckling of grazing dairy cows had a negative effect on machine-milked milk yield, fat percentage and 4% fat-corrected milk yield, but had no effect on udder health or on improved weight gain and body weight at weaning of the calves.     

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.     

Evaluation of specificity of tuberculosis diagnostic assays in caprine flocks under different epidemiological situations

The aim of this study was to evaluate the specificity of the most widely used tuberculosis (TB) diagnostic tests, single intradermal tuberculin (SIT) and single comparative intradermal tuberculin (SCIT) tests and interferon-gamma (IFN-γ) assay in 937 animals from eight TB-free caprine flocks under different epidemiological situations. Maximum specificity was found using SCIT test (99.4-100% depending on the interpretation criteria) while SIT test and IFN-γ assay showed a slightly lower overall specificity (97.6-99.2% and 96.4-98.4% respectively). Specificity of the SIT test in a Corynebacterium pseudotuberculosis infected flock was significantly (P<0.05) lower if a severe interpretation criterion was applied. Similarly, specificity values of SIT test and particularly IFN-γ assay in a paratuberculosis (PTB)-vaccinated flock were lower than those observed in non-vaccinated flocks. Higher proportion of false positive reactors to TB tests (SIT and IFN-γ assay) were observed among animals positive in the PTB-ELISA in PTB vaccinated flock. These results demonstrate that TB diagnostic tests show an adequate specificity when performed in goats from TB-free flocks in most situations. However, certain factors such as C. pseudotuberculosis infection and paratuberculosis vaccination can have a negative impact in the most sensitive tests.     

Comparative pharmacokinetics and a clinical laboratory evaluation of intravenous acetaminophen in Beagle and Galgo Español dogs

Objective

To assess the pharmacokinetics (PK) and conduct a clinical laboratory evaluation of acetaminophen in Beagle and Galgo Español (GE) dogs.

Intestinal bacterial overgrowth includes potential pathogens in the carbohydrate overload models of equine acute laminitis

Carbohydrate overload models of equine acute laminitis are used to study the development of lameness. It is hypothesized that a diet-induced shift in cecal bacterial communities contributes to the development of the pro-inflammatory state that progresses to laminar failure. It is proposed that vasoactive amines, protease activators and endotoxin, all bacterial derived bioactive metabolites, play a role in disease development. Questions regarding the oral bioavailability of many of the bacterial derived bioactive metabolites remain. This study evaluates the possibility that a carbohydrate-induced overgrowth of potentially pathogenic cecal bacteria occurs and that bacterial translocation contributes toward the development of the pro-inflammatory state. Two groups of mixed-breed horses were used, those with laminitis induced by cornstarch (n=6) or oligofructan (n=6) and non-laminitic controls (n=8). Cecal fluid and tissue homogenates of extra-intestinal sites including the laminae were used to enumerate Gram-negative and -positive bacteria. Horses that developed Obel grade2 lameness, revealed a significant overgrowth of potentially pathogenic Gram-positive and Gram-negative intestinal bacteria within the cecal fluid. Although colonization of extra-intestinal sites with potentially pathogenic bacteria was not detected, results of this study indicate that cecal/colonic lymphadenopathy and eosinophilia develop in horses progressing to lameness. It is hypothesized that the pro-inflammatory state in carbohydrate overload models of equine acute laminitis is driven by an immune response to the rapid overgrowth of Gram-positive and Gram-negative cecal bacterial communities in the gut. Further equine research is indicated to study the immunological response, involving the lymphatic system that develops in the model.     

Succession and management of tropical dry forests in the Americas: Review and new perspectives

Understanding tropical forest succession is critical for the development of tropical forest conservation strategies worldwide, given that tropical secondary forests can be considered the forests of the future. Tropical dry forests (TDF) are among the most threatened tropical ecosystems, there are more secondary forests and forest restoration efforts that require a better understanding of successional processes. The main goal of this synthesis for this special issue on the ecology and management of tropical dry forests in the Americas is to present a summarized review of the current knowledge of the ecology and management implications associated to TDF succession. We explore specific issues associated to tropical dry forest succession with emphasis on the use of chronosequences, plant diversity and composition, plant phenology and remote sensing, pollination, and animal–plant interactions; all under the integrating umbrella of ecosystem succession. We also emphasize the need to conduct socio-ecological research to understand changes in land-use history and its effects on succession and forest regeneration of TDF. We close this paper with some thoughts and ideas associated with the strong need for an integrating dimension not considered until today: the role of cyberinfrastructure and eco-informatics as a tool to support sound conservation, management and understanding of TDF in the Americas.