Degradation of Cry1Ab protein from genetically modified maize in the bovine gastrointestinal tract

Immunoblotting assays using commercial antibodies were established to investigate the unexpected persistence of the immunoactive Cry1Ab protein in the bovine gastrointestinal tract (GIT) previously suggested by enzyme-linked immunosorbent assay (ELISA). Samples of two different feeding experiments in cattle were analyzed with both ELISA and immunoblotting methods. Whereas results obtained by ELISA suggested that the concentration of the Cry1Ab protein increased during the GIT passage, the immunoblotting assays revealed a significant degradation of the protein in the bovine GIT. Samples showing a positive signal in the ELISA consisted of fragmented Cry1Ab protein of approximately 17 and 34 kDa size. Two independent sets of gastrointestinal samples revealed the apparent discrepancy between the results obtained by ELISA and immunoblotting, suggesting that the antibody used in the ELISA reacts with fragmented yet immunoactive epitopes of the Cry1Ab protein. It was concluded that Cry1Ab protein is degraded during digestion in cattle. To avoid misinterpretation, samples tested positive for Cry1Ab protein by ELISA should be reassessed by another technique.     

Coherent response of a group of plantation-grown Scots pine trees to wind loading

Wind-induced motion of 29 neighbouring trees growing in a Scots pine plantation was measured over the period 14 March to 24 March 2008. The bi-orthogonal decomposition (BOD) of the complex tree motion field into a limited number of spatio-temporal modes provided the basis for the analysis of the response behaviour to wind excitation of the group of sample trees. It is shown that the first BOD-mode was the most energetic and differed from all other BOD-modes. The BOD-results as well as the results from wavelet analysis of the temporal eigenvectors of the BOD-modes demonstrate that two concomitant low-frequency components in the streamwise wind velocity component stimulated coherent response of the sample trees at the tree group level. However, in the range of the wind speed measured close to the top of the Scots pine forest canopy (hourly mean wind speed values lower than 6 m s⁻¹), the wind loads associated with these low-frequency airflow structures were too low to harmonise the motion of the sample trees completely. It is shown that instantaneous single tree responses to wind excitation were highly irregular in magnitude and direction. Results from Fourier and wavelet analysis demonstrate that sway in the first mode dominated the wind-induced sway behaviour at the tree level.     

Effects of region and cultivar on alkylresorcinols content and composition in wheat bran and their antioxidant activity

This study evaluated the effects of cultivar and region on the composition of alkylresorcinols (ARs) of 24 wheat bran samples from 6 cultivars grown in four locations (Bath, Nairn, Palmerstone and Ridgetown) in Ontario (ON), Canada, using gas chromatography/mass spectrometry (GC/MS). Total phenolic content (TPC) of wheat bran extracts was determined by the Folin–Ciocalteau method and the antioxidant activity of wheat bran extracts was measured by 2, 2-Dipheny-1-picryhydrazyl radical (DPPH) scavenging activity and oxygen radical absorbance capacity (ORAC). The highest ARs content (μg/g) was found in cultivars Emmit (1522), Harvard (1305), Warthog (1170), and Superior (853), grown in Ridgetown. The relative saturated and unsaturated ARs (%) were 89 and 11, respectively. Total ARs content, their composition, TPC and antioxidant activity of wheat bran extracts were significantly (P < 0.05) affected by location and cultivar and their interaction. TPC, %DPPH_dis, and ORAC values for different wheat bran sample extracts ranged from 3 to 58 (mg FAE/g), 5 to 68 (%), and 6 to 94 (μmol TE/g), respectively. Our work provides a detailed examination of region and cultivar effects on potential of ARs in wheat bran and the results can be used for screening and breeding purposes.     

Evaluation of a 15‐week CHOP protocol for the treatment of canine multicentric lymphoma

Dose intense CHOP protocols have been shown to improve outcome for people with non‐Hodgkin's lymphoma, but evaluation of dose intense CHOP protocols for canine lymphoma is currently limited. The hypothesis of this retrospective study was that a 15‐week dose intense CHOP protocol would have shorter treatment duration with similar efficacy to other doxorubicin‐based multidrug protocols. Thirty‐one client owned dogs with multicentric lymphoma were treated with a 15‐week CHOP chemotherapy protocol with an overall response rate of 100% and a median progression‐free interval (PFI) of 140 days [95% confidence interval (CI) 91–335 days]. Dogs that had two or more treatment delays had significantly prolonged PFI and overall survival in multivariate analysis. Dose intensity did not correlate with patient outcome. Dogs experiencing multiple treatment delays secondary to adverse events may receive their individual maximally tolerated dose while dogs with no adverse events may be underdosed. Future studies should focus on individual patient dose optimization.     

Rhizodeposition of C and N in peas and oats after C-N double labelling under field conditions

Compounds released by plant roots during growth can make up a high proportion of below-ground plant (BGP) carbon and nitrogen, and therefore influence soil organic matter turnover and plant nutrient availability by stimulating the soil microorganisms. The present study was conducted to examine the amount and fate of C (CdfR) and N rhizodeposits (NdfR), in this study defined as root-derived C or N present in the soil after removal of roots and root fragments, released during reproductive growth. BGP biomass of peas (Pisum sativum L.) and oats (Avena sativa L.) was successfully labelled in situ with a ¹³C-glucose-¹⁵N-urea mixture under field conditions using a stem feeding method. Pea plants were labelled at the beginning of flowering and harvested 36 and 52 days after labelling at pod filling (P_P) and maturity (P_M), respectively. Oat plants were labelled at grain filling and harvested 42 days after labelling at maturity (O_M). CdfR was 24.2% (P_P), 29.6% (P_M) and 30.8% (O_M) of total recovered plant C. NdfR was 32.1% (P_P), 36.4% (P_M) and 30.0% (O_M) of total plant N. Due to higher N assimilation, amounts of NdfR were four times higher in peas in comparison with oats. The results for NdfR in peas were higher than results from other studies. The C-to-N ratio of rhizodeposits was lower under peas (17.3) than under oats (41.9) at maturity. At maturity, microbial CdfR at 0-30 cm soil depth was 37% of the microbial biomass C in peas and 59% in oats. Microbial NdfR was 15% of microbial N in peas and 5% in oats. Furthermore, inorganic NdfR was 34% in peas and 9% in oats at 0-30 cm at maturity. These results show that rhizodeposits of peas provide a more easily available substrate to soil microorganisms, which are incorporated to a greater extent and turned over faster in comparison with oats. Beside the higher amounts of N released from pea roots, this process contributes to the higher N-availability for subsequent crops.     

Effects of temperature on the final stages of sexual maturation in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.)

Maturing male and female Atlantic salmon (Salmo salar L.) were held under three temperature regimes for 10 weeks between September and December: warm (constant 14–16 °C), ambient (decreasing from 11 to 5 °C), and cold (decreasing from 7 to 3 °C). Blood samples were analyzed for plasma steroid levels, and the fish were inspected for the presence of expressible milt (total volume and spermatocrit) and ovulation weekly. Samples of eggs were dry-fertilized with milt stripped from three males held at the same temperatures and incubated until the eyed stage. In females, levels of plasma testosterone (T) and 17β-oestradiol (E₂) dropped as ovulation approached, concurrent with a rapid increase in levels of plasma 17α,20β-dihydroxy-4-pregnen-3-one (17,20β-P). In males, levels of T and 11-ketotestosterone (11-KT) peaked 2–3 weeks after the first appearance of expressible milt, while levels of 17,20β-P increased steadily and did not exhibit a definite peak. Exposure of females to cold water amplified and advanced the profiles of all three steroids compared with the ambient group, and increased the survival rates to the eyed egg stage. Cold water had no immediate effect on the male steroid profiles, but later, higher levels of 17,20β-P were evident compared with both the ambient controls and the warm water group, while the effects on 11-KT and T were more variable. Exposure to warm water completely inhibited both milt production and ovulation. Moreover, warm water modulated the steroid profiles of the males with lower 11-KT levels compared with ambient controls and lower 17,20β-P level compared with cold-water-treated males. In females, warm water resulted in total inhibition of the peri-ovulatory peak in 17,20β-P and prevented the normal decline of T and E₂ levels associated with ovulation. The findings of the present study are highly relevant for broodstock management in aquaculture, as well in understanding the impact of climate change/temperature variability on wild salmon spawning.     

Shearing male lambs in the cold season improves the carcass yield without affecting fattening performance

This research was carried out to determine the effects of shearing of different genotypes of male lambs (one from pure Karayaka stock and two from its crossbreed) in winter on bodyweight gain, feed consumtion, feed conversion efficiency, rectal temperature and carcass yield. A total of 21 lambs aged 8 months were allocated randomly to experimental groups according to a 2 × 3 factorial arrangement for shorn and unshorn animals. The lambs were fed grass hay (100 g/lamb/day) and commercial concentrate feed ad libitum during a 54‐day of fattening period. The lambs were shorn in the middle of the experiment (27 days after beginning the trial). The interaction between the shearing treatment × genotype of the studied parameters was found to be insignificant. The shearing process increased the rectal temperature (P < 0.01), hot carcass yield (P < 0.05) and dressing percentage (P < 0.05) and decreased the weight of the edible inner organs (P < 0.01). Some of these parameters were affected by the genotypes. It was concluded that shearing male lambs in the winter can have a beneficial effect on the hot carcass weight and dressing percentage without affecting fattening performance, and the performance of Karayaka lambs and its crossbred male lambs were similar under the conditions of the present study.     

Impact of growing maize (<Emphasis Type="Italic">Zea mays</Emphasis>) on the decomposition of incorporated fresh alfalfa residues

In this study, the effects of growing maize plants on the microbial decomposition of easily degradable plant residues were investigated in a 90-day pot experiment using a sandy arable soil. Four treatments were carried out: (1) untreated control, (2) with freshly chopped alfalfa residues (Medicago sativa L.) incorporated into soil, (3) with growing maize plants (Zea mays L.), and (4) with growing maize plants and freshly chopped alfalfa residues incorporated into soil. The amount of alfalfa residues was equivalent to 1.5 mg C g⁻¹ soil and 120 μg N g⁻¹ soil. At the end of the experiment, only the combination of growing maize plants and alfalfa residues significantly increased the contents of microbial biomass C, microbial biomass N, and ergosterol in soil compared to the non-amended control. The dry weight of the maize shoot material was more than doubled in the treatment with alfalfa residues than without. In treatment (2), 6% of the alfalfa residues could be recovered as plant remains >2 mm. In treatment (4), this fraction contained 14.7% alfalfa residues and 85.3% maize root remains, calculated on the basis of δ ¹³C values. This means that 60% more alfalfa-C was recovered than in treatment (2). The reasons for the retardation in the breakdown of alfalfa residues might be water deficiency of soil microorganisms in the increased presence of maize roots. Assuming that the addition of alfalfa residues did not affect the decomposition of native soil organic matter, only 23% of the alfalfa residues were found as CO₂ monitored with a portable gas analyzer with a dynamic chamber. The discrepancy is probably due to problems in measuring peak concentrations of CO₂ evolution in the two alfalfa treatments at the beginning of the experiment and in the two maize treatments at the end, especially in treatment (4).     

Efficiency of Two Enucleation Methods Connected to Handmade Cloning to Produce Transgenic Porcine Embryos

The purpose of our work was to establish an efficient-oriented enucleation method to produce transgenic embryos with handmade cloning (HMC). After 41–42 h oocytes maturation, the oocytes were further cultured with or without 0.4 μg/ml demecolcine for 45 min [chemically assisted handmade enucleation (CAHE) group vs polar body (PB) oriented handmade enucleation (OHE) group respectively]. After removal of the cumulus cells and partial digestion of the zona pellucida, oocytes with visible extrusion cones and/or polar bodies attached to the surface were subjected to oriented bisection. Putative cytoplasts without extrusion cones or PB were selected as recipients. Two cytoplasts were electrofused with one transgenic fibroblasts expressing green fluorescent protein (GFP), while non-transgenic fibroblasts were used as controls. Reconstructed embryos were cultured in Well of Wells (WOWs) with porcine zygote medium 3 (PZM-3) after activation. Cleavage and blastocyst rates were registered on day 2 and day 7 of in vitro culture respectively. Meanwhile, the total blastocyst cell number was counted on day 7. We found that the difference was only observed between blastocyst rates (38.6 ± 2% vs 48.1 ± 3%) of cloned embryos with GFP transgenic fibroblast cells after CAHE vs OHE. With adjusted time-lapse for zonae-free cloned embryos cultured in WOWs with PZM-3, it was obvious that in vitro developmental competence after CAHE was compromised when compared with the OHE method. OHE enucleation method seems to be a potential superior alternative method used for somatic cell nuclear transfer (SCNT) with transgenic fibroblast cells.