Novel purification method for recombinant 3AB1 nonstructural protein of foot-and-mouth disease virus for use in differentiation between infected and vaccinated animals.

An indirect enzyme-linked immunosorbent assay (ELISA) was developed for differentiation of animals infected with foot-and-mouth disease virus (FMDV) from vaccinated animals. The test was based on a highly pure and concentrated preparation of recombinant 3AB1 protein obtained by expression in a prokaryotic system, protein separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and electro elution. Experimental- and field-serum samples from naive, vaccinated, and infected cattle were tested for anti3AB1 antibody using the ELISA. A cutoff level was set at 35% of the maximum absorbance obtained with a positive control serum (FMDV-infected animal, 21 days postinfection [dpi]). This assay could detect antibodies from sera of animals experimentally infected by contact (n = 118) with a sensitivity of 97.5%. The specificity was 100%, based on negative test results obtained on 109 sera from naive animals. Remarkably, all sera from animals vaccinated either once (n = 102) or twice (n = 30) were negative. In addition, this 3AB1-ELISA could detect seroconversion at 7 dpi in animals inoculated intradermolingually. This assay constitutes an important tool for the rapid detection of FMDV outbreaks in a vaccinated population. In addition, it presents a reliable, economical, and simple method for testing large numbers of serum samples.     

Delineation of specific management areas for coffee cultivation based on the soil–relief relationship and numerical classification

Predicting and mapping productivity areas allows crop producers to improve their planning of agricultural activities. The primary aims of this work were the identification and mapping of specific management areas allowing coffee bean quality to be predicted from soil attributes and their relationships to relief. The study area was located in the Southeast of the Minas Gerais state, Brazil. A grid containing a total of 145 uniformly spaced nodes 50 m apart was established over an area of 31.7 ha from which samples were collected at depths of 0.00–0.20 m in order to determine physical and chemical attributes of the soil. These data were analysed in conjunction with plant attributes including production, proportion of beans retained by different sieves and drink quality. The results of principal component analysis (PCA) in combination with geostatistical data showed the attributes clay content and available iron to be the best choices for identifying four crop production environments. Environment A, which exhibited high clay and available iron contents, and low pH and base saturation, was that providing the highest yield (30.4l ha^?1) and best coffee beverage quality (61 sacks ha^?1). Based on the results, we believe that multivariate analysis, geostatistics and the soil–relief relationships contained in the digital elevation model (DEM) can be effectively used in combination for the hybrid mapping of areas of varying suitability for coffee production.     

Peripheral nerve stimulation under ultrasonographic control to determine the needle‐to‐nerve relationship

ObjectiveTo determine the needle‐to‐nerve distances during electrical nerve location in dogs at different currents and pulse duration using a peripheral nerve stimulator (PNS) under ultrasound control (US), and the minimal electrical thresholds (MET) necessary to obtain a motor response (MR) after achieving needle‐to‐nerve contact.Study designProspective in vivo experimental trial in a clinical settingAnimalsThirty dogs, scheduled for locoregional anaesthesia of the sciatic nerve.MethodsNeedle‐to‐nerve distance was measured ultrasonographically after obtaining the MR of sciatic nerve with 2, 1 and 0.5 mA and pulse duration 0.1 ms (NS0.1). Thereafter the needle was placed in contact with the nerve and MET was determined. The procedure was repeated with 0.3 ms (NS0.3). Finally the needle was reintroduced to contact the sciatic nerve guided only by US, thus MET‐US was determined. Data were analysed using Kruskal–Wallis or Mann–Whitney tests.ResultsNeedle‐to‐nerve distances were greater when MR was obtained with 2 mA than with 1 and 0.5 mA at 0.1 and 0.3 ms. No significant differences were observed between the needle‐to‐nerve distances using 0.1 or 0.3 ms.The MET [median (range)] was 0.4 (0.18–1.3) mA in NS0.1, 0.32 (0.12–0.8) mA in NS0.3; while MET‐US was 0.7 (0.32–1.5) mA. When the needle contacted the nerve, the MR achieved with currents below 0.3 mA was obtained in 17.2, 40 and 0% of cases using NS0.1, NS0.3 and US respectively.Conclusions and clinical relevanceThe electrical current necessary to obtain a MR decreased as the needle moved towards the nerve. However when the needle tip contacted the nerve, an MR with low current intensity could not be obtained. Thus the absence of motor response at currents below 0.3 mA cannot rule out needle‐epineurium contact. When ultrasound is combined with PNS, it is more important to assess the correct needle position than searching for an MR at low currents.     

Population dynamics of Dasiops inedulis (Diptera: Lonchaeidae) and its biotic and abiotic mortality factors in Colombian sweet passionfruit orchards

Throughout South America, the lonchaeid flies Dasiops spp. are important herbivores of passionfruit crops. However, little is known on the biology and ecology of these insects, resulting in inadequate pest management schemes. In this study, we describe Dasiops inedulis population dynamics in Colombian sweet passionfruit (SP; Passiflora ligularis Juss.) and elucidate biotic mortality factors at different fly developmental stages. From August 2009 to July 2010, D. inedulis and Dasiops spp. abundance was assessed through monthly McPhail bait trapping and collection of SP flower buds, flowers, and immature fruits. Mortality levels of D. inedulis were determined for early instar larvae by ovary dissection and for late-instar larvae or pupae by prey removal trials. Maximum infestation reached 80 % in fruits and flower buds, and bud infestation correlated with precipitation during the previous month. Two days after oviposition, 8.2 ± 2.3 (mean ± SD) Dasiops sp. eggs were found in SP ovaries and 4.4 ± 1.2 late-instar larvae were recovered from immature fruits at day 14. Upon larval drop on the orchard soil, 74.8 % larvae burrowed within the soil within 9 min, while 12.5 % larvae were attacked by ants. In-field mortality of young pupae amounted to 75.3 ± 7.0 %, with vertebrate predators likely causing 12.1 ± 6.0 % mortality. Late-instar larvae and pupae appear highly vulnerable to natural enemy action, with the ground-foraging predator community mainly composed of ants (80.37 %) and ground beetles (9.17 %). Our findings should help develop integrated pest management (IPM) tactics for SP crops.     

Effects of cryopreservation on the mitochondrial bioenergetics of bovine sperm

This study evaluated the bioenergetic map of mitochondria metabolism in cryopreserved bovine sperm. The detected oligomycin-sensitive basal respiration supported ATP production; frozen–thawed spermatozoa were found to have a coupling efficiency higher than 0.80. Cell respiration, however, was not stimulated by the protonophoric action of FCCP, as its titration with 1, 2, 4 and 6 μM did not stimulate the uncoupling activity on oxidative phosphorylation as highlighted by unresponsive oxygen consumption. The unusual effect on the stimulation of maximal respiration was not related to fibronectin- or PDL-coated plates used for cellular metabolism analysis. Conversely, irradiation of frozen–thawed bovine sperm with the red light improved mitochondrial parameters. In effect, the maximal respiration of red-light-stimulated sperm in PDL-coated plates was higher than the non-irradiated. In spite of this, red-light irradiation had no impact on membrane integrity and mitochondrial activity evaluated by epifluorescence microscopy.