Comparison of analytical methodologies based on 1h and 31p NMR spectroscopy with conventional methods of analysis for the determination of some olive oil constituents

The present study was designed to assess the agreement between analytical methodologies based on 1H and 31P NMR spectroscopy and conventional analytical methods (titration, gas chromatography, and high performance liquid chromatography) for measuring certain minor and major constituents (free acidity, fatty acids, iodine value, and phenolic compounds) of olive oil. The standard deviations of the NMR method were comparable to those of the conventional methods, except perhaps those of the total hydroxytyrosol and total tyrosol. Linear regression analyses showed strong correlations between NMR and conventional methods for free acidity, total hydroxytyrosol, total tyrosol, total diacylglycerols, (+)-pinoresinol, (+)-1-acetoxypinoresinol, and apigenin; good correlations for linoleic acid, free hydroxytyrosol, and free tyrosol; and weak correlations for oleic acid, linolenic acid, saturated fatty acids, and luteolin. Furthermore, a method comparison study was conducted and the agreement between NMR and conventional methods was evaluated by using the Bland and Altman statistical analysis. The distribution of the data points in the bias plot showed that 96.4% and 100% of the measurements of free acidity and iodine value, respectively, were within the limits of agreement of the two methods. For the remaining constituents of olive oil, the percentage of measurements, located within the limits of agreement, ranged from 94% to 98.5%.     

Structure-DPPH* scavenging activity relationships: parallel study of catechol and guaiacol acid derivatives

The scavenging behavior of a series of catechol and guaiacol acid derivatives toward DPPH(*) was examined having as a starting point the order of activity derived on the basis of theoretically calculated BDE values. The studied compounds were protocatechuic, homoprotocatechuic, dihydrocaffeic, and caffeic acids and also vanillic, homovanillic, dihydroferulic, and ferulic acids. Catechol and guaiacol were used as reference compounds. Observations from the parallel study were made with regard to structural features (number and position of hydroxyl groups and the side-carbon chain characteristics) that regulated the behavior of the compounds experimentally. The exceptional DPPH(*) scavenging behavior observed for homoprotocatechuic acid in ethanol and for caffeic acid in acetonitrile could not be supported by the respective BDE values. Ferulic was the most active among guaiacolic acids, whereas dihydroferulic exhibited the highest stoichiometry. Ionizable carboxylic groups seem to affect considerably the relative order of activity as was also evidenced using the ORAC assay. Questions raised about the validity of widely accepted views on criteria for SARs are discussed with regard to literature findings.     

Kinetics of maneb degradation during thermal treatment of tomatoes

The kinetics of maneb degradation in tomato homogenates at high temperatures and at two pH values (4 and 9) and the rate of formation of the toxic metabolite, ethylenethiourea (ETU), were studied. Maneb was measured as carbon disulfide by headspace gas-chromatography and ETU by high-performance liquid chromatography with photodiode array detection. First-order kinetics adequately described the degradation of maneb in tomato homogenates. The degradation rate constants exhibited an Arrhenius temperature dependence in the range from 50 to 90 degrees C and the apparent activation energy (E(a)) was calculated to be 36 KJ mol(-1) in homogenates with natural pH (4). Raising temperature from 60 to 75 and to 90 degrees C, ETU formation was significantly increased. Interestingly, the selectivity toward ETU showed a downward trend when the total conversion increased at longer heating times. When the pH of the tomato homogenates was adjusted to 9, the degradation of maneb proceeded faster at both 60 and 90 degrees C. The combination of alkaline pH and the highest temperature (90 degrees C) resulted in the maximum ETU conversion rates. The results of the present study on the fate of maneb and ETU residues during tomato processing, may prove valuable in estimating potential risk from dietary exposure.     

Ozone deposition onto bare soil: A new parameterisation

The variables controlling ozone deposition onto bare soil are still unknown and it is necessary to understand this pathway well, as it represents a significant sink for ozone. Eddy-covariance measurements of ozone (O₃) fluxes were performed over bare soils in agricultural land. Three datasets with contrasted meteorological conditions and soil nitric oxide (NO) emissions were used to study the factors controlling soil deposition. It is considered that ozone deposition can be represented with an aerodynamic resistance (R_a), a quasi-laminar boundary layer resistance (Rb O₃), and an additional resistance, named soil resistance (R_soil). Although it is assumed in previous studies that soil resistance is a function of soil water content (SWC) and could be considered constant as variation of SWC at monthly scale are generally weak, the results of this study indicate that SWC is not the main factor controlling R_soil which shows daily and hourly variations. The main factor controlling soil resistance is the surface relative humidity which is positively correlated with R_soil, contrary to non stomatal resistance onto canopies which show a negative correlation with relative humidity. The relationship between R_soil and the surface relative humidity is probably due to a decrease in the surface available for ozone deposition, due to an increasing adsorption of water molecules onto the ground with relative humidity. A new parameterisation of R_soil was established, where R_soil is a function of the surface relative humidity only (R_soil = R_{soil min} × e^(k×RH_surf), and R_{soil min} = 21 ± 1.01 s m⁻¹ and k = 0.024 ± 0.001, mean ± SD). The measured and parameterised ozone deposition velocities agree well when soil NO emissions are negligible. However, when there are large soil NO emissions, the parameterised ozone deposition strongly underestimates the measured deposition velocity even if the chemical destruction of ozone by reaction with NO in the air column was evaluated to be negligible. This suggests that soil NO emissions enhance soil ozone deposition by chemical reaction at or near the soil surface. The new parameterisation allows a better estimation of soil deposition, especially during daytime when R_soil is overestimated using previously published parameterisations. It is an important step towards a better parameterisation of the non-stomatal uptake of ozone.     

Duplex real-time polymerase chain reaction for simultaneous detection and quantification of Anaplasma marginale and Anaplasma centrale

Anaplasma marginale and Anaplasma centrale are rickettsial pathogens responsible for acute disease and mild infections, respectively, in cattle herds. A duplex real-time polymerase chain reaction (PCR) assay with probes labeled with different fluorophores was developed for simultaneous detection and quantification of A. marginale and A. centrale DNA in bovine blood samples. The assay was able to detect as few as 10(1) and 10(2) DNA copies for A. marginale and A. centrale, respectively, with optimal specificity and reproducibility. Analysis by real-time and nested PCR carried out on 54 samples previously tested by reverse line blot hybridization showed that the established duplex real-time PCR assay can detect and quantify the 2 Anaplasma spp., even if present simultaneously in the same blood samples. Such an assay could be used in pathogenesis studies on bovine acute anaplasmosis.     

Risk factor analysis for antibodies to Brucella, Leptospira and C. burnetii among cattle in the Adamawa Region of Cameroon: a cross-sectional study

Brucellosis, leptospirosis and Q fever are important livestock diseases, commonly responsible for significant production losses, yet their epidemiology in sub-Saharan Africa is largely unknown. Animal reservoirs pose the main risk of transmission to humans, where serious disease can occur. In the developing world setting, the flu-like symptoms of the acute stages of these diseases can be misdiagnosed as malaria, which can result in the administration of the wrong treatment, prolonged disease and increase in antibiotic resistance. Multivariable mixed-effects logistic regression models in this study revealed potential risk factors associated with the aforementioned pathogens in cattle in the Adamawa Region of Cameroon, with wildlife, namely, buffaloes, playing a major role in both Brucella and Coxiella burnetii seropositivity. Cattle mixing with other herds at night and cattle grazing in an area on a route taken by herds on transhumance appear to be positively associated with Leptospira seropositivity, while female cows and whether buffaloes are seen during grazing or transhumance are positively associated with C. burnetii seropositivity. On the other hand, animals that have been on transhumance in the past year and animals belonging to herdsmen of the Fulbe ethnic group appear to be protected against Leptospira and C. burnetii, respectively. Cattle of more than 2 years old appear to have increased odds of being seropositive to either pathogen. Further research is needed to confirm these findings and improve the knowledge of the epidemiology of these three pathogens in Africa, taking particular consideration of the wildlife involvement in the disease transmission.     

Genotypic variation in drought response of silver birch (Betula pendula): leaf water status and carbon gain

To assess genotypic variation in drought response of silver birch (Betula pendula Roth), we studied the plasticity of 16 physiological traits in response to a 12-14-week summer drought imposed on four clones in two consecutive years. In a common garden experiment, 1-year-old clonal trees from regions with low (550 mm year(-1)) to high rainfall (1270 mm year(-1)) were grown in 45-l pots, and leaf gas exchange parameters, leaf water potentials, leaf osmotic potentials and leaf carbon isotope signatures were repeatedly measured. There were no clonal differences in leaf water potential, but stomatal conductance (gs), net photosynthesis at ambient carbon dioxide concentration, photosynthetic water-use efficiency, leaf carbon isotope composition (delta13C) and leaf osmotic potentials at saturation (Pi0) and at incipient plasmolysis (Pip) were markedly influenced by genotype, especially gs and osmotic adjustment. Genotypes of low-rainfall origin displayed larger osmotic adjustment than genotypes of high-rainfall origin, although their Pi0 and Pip values were similar or higher with ample water supply. Genotypes of low-rainfall origin had higher gs than genotypes of high-rainfall origin under both ample and limited water supply, indicating a higher water consumption that might increase competitiveness in drought-prone habitats. Although most parameters tested were significantly influenced by genotype and treatment, the genotype x treatment interactions were not significant. The genotypes differed in plasticity of the tested parameters and in their apparent adaptation to drought; however, among genotypes, physiological plasticity and drought adaptation were not related to each other. Reduction of gs was the first and most plastic response to drought in all genotypes, and allowed the maintenance of high predawn leaf water potentials during the drought. None of the clones exhibited non-stomatal limitation of photosynthesis. Leaf gs, photosynthetic capacity, magnitude of osmotic adjustment and delta13C were all markedly lower in 2000 than in 1999, indicating root limitation in the containers in the second year.