Comparative activity of Choristoneura fumiferana nucleopolyhedrovirus propagated in different hosts

The biological activity of the Ireland strain of Choristoneura fumiferana (Clem) nucleopolyhedrovirus (CfMNPV) propagated in different hosts was determined to provide the basis upon which genetically modified CfMNPV, or other naturally occurring isolates, should be compared. Occlusion bodies (OB) derived from CF-203 cells were significantly larger and more pathogenic than those propagated in vivo when tested against the fifth larval instar of C fumiferana (Clem) and C occidentalis Freeman. The dose-responses (LD50 and LD95, expressed as occlusion bodies per larva) of C fumiferana larvae to in vitro-propagated OBs were 274 and 5785, respectively. The values of LD50 and LD95 to C occidentalis larvae were 19 and 118, respectively. There were no significant differences in pathogenicity or size when OBs propagated in C fumiferana larvae were tested against either insect species, nor were there significant differences for OBs propagated in C occidentalis larvae. The LD50 and LD95 of in vivo-produced OBs to C fumiferana were 925 and 61988, respectively. The LD50 and LD95 to C occidentalis were 50 and 453, respectively. OBs propagated in vitro had a mean volume of 13.13 microm3, whereas those propagated in vivo ranged from 0.84 to 1.41 microm3. The median survival time-responses (ST50) of fifth-instar C fumiferana or C occidentalis larvae to OBs propagated in vivo were not significantly different from those propagated in vitro at the dosage levels tested. Values of ST50 of C fumiferana larvae to in vitro- and in vivo-produced OBs at dosages causing less than 50% mortality rangedfrom 9.6 to 9.8 days post-inoculation (dpi), whereas a LD95 dose resulted in ST50 values ranging from 7.3 to 7.7 days. ST50 values of C occidentalis larvae at dosages causing less than 50% mortality ranged from 9.8 to 10.2 dpi, whereas a LD95 dose resulted in ST50 values ranging from 9.5 to 9.8 dpi. The median feeding cessation time-response (FT50) of fifth-instar C fumiferana larvae to OBs propagated in vitro (5.7 days) was not significantly different from the FT50 of those propagated in vivo in either insect species (5.3 and 5.7 days) at the dosage level tested (LD95). No significant differences in FT50 values were observed between OBs propagated in either larval host. The FT50 of C occidentalis larvae to OBs propagated in vitro (7.7 days) was not significantly different from that to those propagated in vivo in C occidentalis larvae (7.6days), but somewhat different (7.2 days) from that to those propagated in C fumiferana larvae. Results indicate that CfMNPV can be propagated in vivo in either C fumiferana or C occidentalis larvae (or sequentially through both) without alteration in infectivity, although the use of the CF-203 cell line yields the most biologically active OBs.     

DNA hybridization assay for detection of nucleopolyhedrovirus in whitemarked tussock moth (Orgyia leucostigma) larvae

DNA dot‐blot hybridization assays utilizing a horseradish peroxidase‐labelled whole genomic DNA probe and enhanced chemiluminescence were conducted to quantify detection thresholds of nucleopolyhedrovirus (NPV) in whitemarked tussock moth (Orgyia leucostigma) larvae. The minimum detection thresholds for an aqueous suspension of occlusion bodies (OBs), OBs added to macerates of non‐infected larvae and OBs in macerates of diseased larvae were 7.8 × 10³, 7.8 × 10³, and 1.5 × 10³ OBs, respectively. Purified viral DNA was detected at a concentration of 1.6 × 10⁻¹ ng in a 20 µl volume. The presence of pre‐occluded viral nucleocapsids and DNA, inherent to infected larvae, improved the detection threshold five‐fold compared with OBs alone. Larval tissues did not block the detection system utilized, nor did they bind non‐specifically to the probe. Detection thresholds, upon sequential hybridization of the same membrane, on average deteriorated two‐fold between the first and second hybridization and an additional six‐fold between the second and third hybridization. NPV infection was detected two days post‐inoculation (pi) in about one‐third of the larvae examined and in almost all larvae three days pi. Microscopic analysis of stained larval smears missed NPV infection in almost all larvae two days pi and about two‐thirds of the larvae three days pi. Results from the two methods of analysis were not comparable until four days pi. The detection system utilized is a reliable, efficient and simple method for the early detection of NPV infection in large numbers of larvae and may be used for further studies quantifying the role of this baculovirus in the ecology of whitemarked tussock moth populations. © 2001 Society of Chemical Industry     

Preparation of Highly Efficient CoFe<Subscript>2</Subscript>O<Subscript>4</Subscript>/Zn<Subscript>2</Subscript>SnO<Subscript>4</Subscript> Composite Photocatalyst for the Degradation of Rhodamine B Dye from Aqueous Solution

CoFe₂O₄/Zn₂SnO₄ composite was synthesized using a simple two-step process and applied as a novel-efficient photocatalyst for the rhodamine B degradation from aqueous solution. Characterization techniques such as X-ray diffraction (XRD), N₂ adsorption-desorption isotherms, scanning electron microscopy (SEM), EDS analysis, and diffuse reflectance spectroscopy were employed in order to investigate the physical and chemical properties of composite. Higher values of the specific surface area, pore volume and diameter, and a smaller band-gap energy promoted a greater catalytic activity of CoFe₂O₄/Zn₂SnO₄ composite when compared to Zn₂SnO₄. A rapid decolorization of dye solution was observed at 40 min of reaction using the CoFe₂O₄/Zn₂SnO₄ catalyst, being 2.5 times faster than the Zn₂SnO₄ alone. Therefore, the CoFe₂O₄/Zn₂SnO₄ composite shows extraordinarily high photocatalytic activity toward the degradation of rhodamine B dye from aqueous solution.     

Antinociceptive effects of tramadol and acepromazine in cats

Effects of tramadol and acepromazine on pressure and thermal thresholds were examined in eight cats. After baseline measurements, subcutaneous (SC) tramadol 1 mg/kg, acepromazine 0.1 mg/kg, tramadol 1 mg/kg with acepromazine 0.1 mg/kg, or saline 0.3 ml were given. Serial measurements were made for 24 h. Mean thermal thresholds did not change significantly [analysis of variance (ANOVA)] from baseline. The maximum thermal threshold increase above baseline was 2.8+/-2.8 degrees C at 6 h (P>0.05) after tramadol; it was above the 95% confidence interval (CI) at 0.75, 3 and 6 h. Pressure thresholds increased above baseline from 0.25 to 2 h after acepromazine (P<0.05) and from 0.5 to 3 h after the combination (P<0.05), with a maximum increase of 132+/-156 mmHg 0.25 h after acepromazine and 197+/-129 mmHg 0.5 h after the combination. Pressure thresholds were above the 95% CI from 0.25 to 2 h after acepromazine and from 0.5 to 3 h after the combination. SC tramadol at 1 mg/kg in cats had limited effect on thermal and pressure nociception, but this was enhanced by acepromazine. Acepromazine alone had pressure antinociceptive effects.     

Alkaline hydrolyzable nitrogen and properties that dictate its distribution in paddy soil profiles

Nitrogen (N) is a key nutrient for rice production, and its bioavailability in paddy soils is strongly coupled to soil organic matter (SOM) cycling. A better understanding of potentially available N forms in soil, such as alkaline hydrolyzable N (AH-N), and their depth distribution will support the development of best management practices to improve the N use efficiency of rice while minimizing adverse environmental effects. Fifteen rice (Oryza sativa L.) fields from Southern Brazil were selected, and stratified soil samples were taken to a depth of 60 cm before crop establishment. Selected soil physical and chemical properties were analyzed to evaluate their relationships with AH-N contents in the soil profile. The AH-N contents below 20 cm varied extensively (increased, reduced, or constant) compared with that above 20 cm. Although clay and clay + silt contents were highly correlated to AH-N for some soils, the major property dictating AH-N distribution by depth was total N (TN), as the correlation between TN and AH-N was mainly by direct effect. The proportion of TN recovered as AH-N across sites and depths presented high amplitude, and thus AH-N was not a constant N pool across depths, indicating that AH-N can be affected by soil management practices even when TN showed no major changes. The distinct distribution of AH-N across soil sampling sites and depths indicates that depths greater than 20 cm should be considered when calibrating the AH-N index for N fertilizer recommendations for flooded rice in Southern Brazil.     

Development of a pressure nociceptive threshold testing device for evaluation of analgesics in cats

A pressure analgesiometric device was developed for unrestrained cats. Eleven cats were studied. Stimulation was via three rounded pins within a bracelet on the forearm. The pins were advanced by manual bladder inflation. Bladder pressure was measured using a strain gauge pressure transducer. The threshold was recorded at the behavioural end point. Thresholds were measured at 5 and 15min intervals for 2-4h, after removal/replacement of the cuff, for 120min after SC butorphanol (0.4mg/kg), and with mild skin inflammation at the testing site. Data were analysed using ANOVA. Pressure thresholds in untreated cats were around 150mmHg. The minimum interval for testing was established as 15min. Data were reproducible over 4h and beyond 24h. Thresholds in 5 cats increased (P<0.05) above baseline for 45min after butorphanol with a maximum increase of 270+/-182mmHg at 10min. Thresholds decreased with inflammation. The method appears suitable for feline analgesia investigations.     

Validation of the Italian version of the UNESP–Botucatu unidimensional composite pain scale for the assessment of postoperative pain in cattle

Objective

The aim of this study was to validate the Italian version of the UNESP–Botucatu Unidimensional Composite Pain Scale (UCPS-IV) for assessing postoperative pain in cattle.

LHRH and beta-endorphin in the hypothalamus of the ram in relation to photoperiod and reproductive activity

The regional distribution and relative concentrations of LHRH and beta-endorphin immunoreactivity in the hypothalamus were compared in sexually active and sexually inactive Soay rams. LHRH was localized principally in the medial basal hypothalamus and median eminence with lower concentrations in the anterior hypothalamus and preoptic area. beta-endorphin immunoreactivity was also found in highest concentrations in these regions but was more widely distributed into the rostral, dorsal and lateral parts of the hypothalamus. LHRH content in the medial basal hypothalamus was significantly lower in the sexually active rams compared to inactive rams while there was no difference in beta-endorphin immunoreactivity content between the two groups. Gel filtration chromatography of hypothalamic extracts revealed that the beta-endorphin immunoreactivity was due to both beta-lipotropin and beta-endorphin; the ratio of beta-endorphin: beta-lipotropin tended to be less in the sexually active rams than in the sexually inactive rams. The results are consistent with the hypothesis that beta-endorphin is a physiologically important endogenous opioid involved in the modulation of LHRH secretion.     

QTL mapping for the cooking time of common beans

The decrease in the per capita consumption of beans has been partially attributed to their lengthy cooking time and the aggregated capital costs of their preparation. The aim of this study was to map microsatellite (SSR) markers linked to quantitative trait loci (QTLs) that govern the cooking time of common beans. An F₂ generation consisting of 140 families was generated from a cross between lines CNFM7875 and Laranja. The cooking time of the F_2:4 and F_2:5 generations was then evaluated, and the latter generation was tested in two environments. The analysis of variance found a significant effect for the interactions between the families (P < 0.01) in both the F_2:4 and F_2:5 generations, as well as for the group analyses performed in the two environments. The experimental coefficient of variation varied from 9.42 to 17.94%. The Pearson’s correlation test indicated no significant association between water absorption and cooking time. The heritability coefficients had values of 0.532 and 0.739 for the F_2:5 families evaluated at the two different locations, and the group analysis of the F_2:5 generation indicated that there was a significant genotype × environment interaction. Of the 105 polymorphic SSRs evaluated, 91 mapped to 12 linkage groups with an estimated map size of 1,303.7 cM. Six significant QTLs were detected in both environments, and the percentage of the phenotypic variation that was explained by these loci ranged from 11.54 to 21.63%. As the genetic control was oligogenic, the identification of QTLs should serve as an optimal starting point for the implementation of a selection program.