Progress towards clonal propagation of Camellia japonica cv. Alba Plena by tissue culture techniques

Summary

Optimal rooting conditions have been determined for shoot cultures of Camellia japonica cv. Alba Plena derived from a 50 year old tree: dipping the base of shoots in 1 g l^?l solution for 15 min, followed by 12 days’ darkness, induced 87% rooting in shoots cultured with Woody Plant Medium (WPM) macronutrients. Halving the auxin concentration or exposure time considerably reduced this rate, and root formation was severely inhibited if an initial dark period for the entire shoot was not used. The type of support (agar or paper bridges) did not significantly affect the rooting percentage or number of roots per rooted shoot, but liquid media induced greater root elongation. No significant differences were observed between the use of WPM and a modified Heller’s medium as regards the rooting percentages achieved, but the number of roots formed was considerably greater with WPM, as was the survival rate after transfer to soil (70% for transfer as against 35% with the modified Heller’s medium). Best survival rates were achieved when shoots were transferred to the acclimatization soil 12 days after auxin treatment, i.e. immediately after the dark period.     

Assessment of s-Triazine Catabolic Potential in Soil Bacterial Isolates Applying atz Genes as Functional Biomarkers

Fluorescence in situ hybridization (FISH) technique and qPCR analyses, targeting atz genes, were applied to detect the presence of simazine-degrading bacteria in an agricultural soil with a history of herbicide application. atzB-targeted bacteria detected by FISH represented 5% of total soil bacteria with potential capability to metabolize the herbicide. The soil natural attenuation capacity was confirmed in soil microcosms by measuring simazine degradation. Moreover, four bacterial strains were isolated from the soil and identified as Acinetobacter lwoffii, Pseudomonas putida, Rhizobium sp. and Pseudomonas sp. The isolates were able to grow using different s-triazine compounds and related metabolites as the sole carbon source. Growth parameters in presence of simazine were calculated using the Gompertz model. Rhizobium sp. showed the highest simazine degradation (71.2%) and mineralization (38.7%) rates, whereas the lowest values were found to A. lwoffii??50.4% of degradation and 22.4% of mineralization. Results from qPCR analyses of atzA, atzB and atzC genes revealed their presence in Rhizobium sp. and A. lwoffii, being atzB and atzC the most abundant functional genes. Rhizobium sp. showed a higher amount of the three biomarkers compared to A. lwoffii: the atzA, atzB and atzC gene copy number per microlitre were, respectively, 10¹, 10² and 10³-fold higher in the former. Therefore the proposed molecular approaches based on the use of atz genes as biomarkers can be considered as useful tools to evaluate the presence and potential capability of degrading-s-triazines soil microorganisms.     

An Exploratory Study of Particulate PAHs in Low-Polluted Urban and Rural Areas of Southwest Spain: Concentrations, Source Assignment, Seasonal Variation and Correlations with Other Air Pollutants

A set of 72?PM₁₀ samples from low-polluted urban and rural locations belonging to the regional air monitoring network of Extremadura (Spain) were collected in a 1?year sampling period. Sample pre-treatment and analytical determination by gas chromatography?Cion trap mass spectrometry were optimised and validated for the analysis of the priority 16 US Environmental Protection Agency polycyclic aromatic hydrocarbons (PAHs). The influence of meteorological conditions (temperature, relative humidity and solar radiation) and other atmospheric pollutants (O₃, NO₂, SO₂, PM₁₀) has been covered in detail and Pearson correlation test were used for this purpose. Spatial distribution of particulate PAHs was evaluated and the comparison with other European sites was also established. Possible emission sources were identified and assigned by using molecular diagnostic criteria.     

Late nitrogen fertilization affects nitrogen remobilization in wheat

A pot experiment with wheat plants was carried out to study how late application of nitrogen (N) fertilizer affects the use of pre‐anthesis N reserves during the grain‐filling period. Increasing doses of N fertilizer were applied (0, 40, and 52 mg N plant^–1), either in two amendments (growth stages GS20 and GS30, according to Zadoks scale) or in three amendments (GS20, GS30, and GS37). The experiment was arranged in a complete randomized three‐block design with 129 plants per treatment. The plants were watered daily, harvested every 2 d between anthesis and maturity, and were separated into roots, leaf sheaths, leaf blades, and ears for further N determination. Grain N concentration improved due to a late N application in GS37 by 14% (higher N dose) and by 7% (further splitting the same N‐fertilizer dose, respectively). The higher the N‐fertilizer dose applied, the greater was the amount of pre‐anthesis reserves in vegetative organs, these reserves became later available for remobilization. Although splitting the same N dose in three amendments did not increase the N reserves, these reserves were more efficiently remobilized allowing an improvement in grain N concentration. The fertilizer management did not change the temporary pattern of N accumulation in the ear, but did induce a change in the amount of N remobilized and in the contribution of each organ (root, leaf sheath, leaf blade) to this remobilization. Late N amendment allowed a greater N availability of leaf blades and ear N reserves (from 20% up to 26% and from 19% up to 22%, respectively) for remobilization towards the grain, decreasing the root contribution from 28% down to 15%, while the contribution of leaf sheaths was maintained around 35% irrespective of the N applied.     

Evaluation of preflooding effects on iron extractability and phytoavailability in highly calcareous soil in containers

Previous pot cropping and laboratory incubation experiments were consistent with field observations showing that temporary flooding before cropping can increase the availability of soil Fe to plants. To study the effect of temporary flooding on changes in soil Fe phytoavailability we used 24 highly calcareous, Fe chlorosis–inducing soils to carry out a pot experiment where peanut and chickpea were successively grown after flooding for 30 d. At the end of the cropping experiment, the preflooded soil samples exhibited higher concentrations of acid oxalate‐, citrate/ascorbate‐ and diethylenetriaminepentacetic acid (DTPA)–extractable Fe (Fe_ox, Fe_ca, and Fe_DTPA, respectively) than the control (nonflooded) samples. Also, Fe_ox and Fe_ca exhibited no change by effect of reflooding of the cropped soils or three wetting–drying cycles in freeze‐dried slurries of soils previously incubated anaerobically for several weeks. Leaf chlorophyll concentration (LCC) in both peanut and chickpea was greatly increased by preflooding. The best predictor for LCC was Fe_ox, followed by Fe_ca and Fe_DTPA. The LCC–soil Fe relationships found suggest that the Fe species extracted by oxalate and citrate/ascorbate from preflooded soils were more phytoavailable than those extracted from control soils. However, the increased phytoavailability of extractable Fe forms was seemingly limited to the first crop (peanut). Flooding dramatically increased Fe_DTPA; however, high Fe_DTPA levels did not result in high LCC values, particularly in the second crop. Therefore, this test is a poor predictor of the severity of Fe chlorosis in preflooded soils.     

Relationships between admissions requirements and pre-clinical and clinical performance in a distributed veterinary curriculum

The purpose of this study was twofold: first, to assess the relationships between knowledge-based admission requirements and pre-clinical and clinical performance in a distributed model of veterinary education that uses problem-based learning as the main instruction method in the first two years of the curriculum; second, to compare pre-clinical and clinical performance with performance on the Program for the Assessment of Veterinary Education Equivalence (PAVE) exam. Admissions data including overall GPA, prerequisite GPA, Graduate Record Examination (GRE) score on the Analytical, Analytical Writing, Quantitative, and Verbal sections), veterinary school performance data (GPA for pre-clinical and clinical years), and performance PAVE (taken at the end of second year) were analyzed for two classes (N = 155, 85.8% women and 14.2% men). Overall GPA, prerequisite GPA, and GRE Quantitative and Analytical scores were the best predictors for pre-clinical (years 1 and 2) performance (R = 0.49, 23.5% of the variance), GRE Analytical score was the best predictor for year 3 (pre-clinical and clinical) performance (R = 0.25, 6.3% of the variance), GRE Quantitative score was the best predictor for PAVE performance (R = 0.27, 7.5% of the variance), and GRE Analytical score was the best predictor for clinical performance (year 4; R = 0.21, 4.4% of the variance). PAVE scores correlated with GRE Quantitative scores (r = 0.27, p <.01) and veterinary school performance, with higher correlations in the pre-clinical years (rs = 0.67-0.36, p < .01), providing evidence of convergent validity for the PAVE exam.     

Oleuropein ameliorates acute colitis in mice

Oleuropein, the major secoiridoid in olive tree leaves, possesses a wide range of health promoting properties. It has recently been shown to exhibit anti-inflammatory activity. We have evaluated the effect of oleuropein on dextran sulfate sodium (DSS)-induced experimental colitis in mice in order to provide insight into its mechanisms of action. Oral administration of oleuropein notably attenuated the extent and severity of acute colitis while reducing neutrophil infiltration; production of NO, IL-1β, IL-6, and TNF-α; expression of iNOS, COX-2, and MMP-9; and the translocation of the NF-κB p65 subunit to the nucleus in colon tissue. In LPS-stimulated peritoneal macrophages, the oleuropein metabolite, hydroxytyrosol, was shown to inhibit NO production, iNOS expression, NF-κB p65 subunit translocation, mRNA expression, and the release of IL-1β, IL-6, and TNF-α. These results suggest that the effect of oleuropein on DSS-induced colitis is associated with a decrease in the production of interleukins and expression of proteins, principally through reduction of NF-κB activation.     

Anti-Neospora caninum antibodies in milk in relation to production losses in dairy cattle

A comprehensive field study was carried out with the following objectives: (a) to assess the usefulness of individual and bulk tank milk analysis for determining Neospora caninum serostatus in individual cows and herds, and (b) to study the associations between N. caninum infection status (based on milk testing), and several productive and reproductive parameters in the animals. Antibodies were detected with a commercially available ELISA test (Bio K 192/5). Analysis of paired serum and milk samples from 1134 lactating cows on 38 farms revealed that 97.6% of the ELISA results were coincident, irrespective of whether serum or milk samples were used. Moreover, multiple linear regression analysis revealed that 86.0% of the variations in ELISA values in milk were due to variations in the serum. The measurement of antibodies in bulk tank milk was a good estimator of the herd level status of N. caninum infection, and enabled detection of infection in 94.7% herds with ≥10.0% seropositive cows and/or in all herds with >4% highly seropositive cows. The odds ratio for abortion in seropositive animals was 9.1 times higher than in seronegative animals. The infection serostatus was also a significant risk factor, as the odds ratio for abortion was even higher (12.0 times) in cows categorized as highly seropositive. ELISA values for the bulk milk from 387 randomly selected herds were negatively associated with average milk production. Moreover, milk production losses mainly occurred on farms categorized as highly positive (i.e. herds with ≥20.0% seropositive cows).     

Culture system and long-term storage of culture media in the in vitro production of bovine embryos

The optimum culture system for in vitro matured and fertilised oocytes still remains to be clarified. Culture media (CM) for mammalian embryos are routinely prepared fresh for use and preserved under refrigeration during one or two weeks. The purposes of this work were (1) to compare the efficiency of a synthetic oviduct fluid (SOF) with two different bovine serum albumin (BSA) concentrations (3 and 8 g/L) for the in vitro production of bovine blastocysts, (2) to test the effect of timing on adding fetal calf serum (FCS) to the SOF, and (3) to evaluate the effects on bovine embryo development of freezing and lyophilisation as procedures for preserving the SOF. Supplementation of SOF with 3 g/L BSA increased Day-7 blastocyst expansion rates (18.3 ± 1.6 vs. 14.4 ± 0.7; P < 0.05), although no differences in hatching rates were found. Addition of FCS to SOFaa (SOF with amino acids) medium supplemented with sodium citrate (SOFaaci) at 48 and at 72 h post-insemination (PI) allowed obtaining higher Day-6 embryo development rates than when FCS was added at 18 or 96 h PI (Day-6 morulae + blastocyst rate: 30.0 ± 1.1, 40.8 ± 1.1, 43.9 ± 2.3 and 39.3 ± 0.5 for FCS addition at 18, 48, 72 and 96 h, respectively). Hatching rates were significantly improved when serum was added at 72 h PI. Finally, both refrigeration and lyophilisation appeared as useful cryopreservation procedures for SOFaaci, although a significant loss of its ability to support embryo development, compared to the control fresh culture medium, was observed.